Search

This work aimed to simplify and improve the process of binding monoclonal antibodies (mAbs) covalently to filter paper for use in dried blood spot sampling, enabling instant capture of protein biomarkers for targeted protein determination. Incorporating the necessary immunocapture sample preparation step in the initial sampling stage saves time and reduces the workload. The biomarker human chorionic gonadotropin (hCG) was used as the model analyte. The antibody‐based paper samplers were prepared by functionalizing paper discs (6 mm) through a simple reaction using divinyl sulfone (DVS). After DVS activation, the paper discs were incubated with E27 hCG mAbs, followed by 0.05% tween/phosphate buffer saline to block the surface. After sample application and drying, the discs only needed to be washed before tryptic digestion and finally analysed on a nanoliquid chromatography–tandem mass spectrometry system. The finished DVS‐mAbs samplers could selectively capture hCG (100 ng/mL) from human serum, with a recovery of 50%. Sample clean‐up reduced the number of identified proteins from 132 to 82 before and after wash, respectively, with a 70% reduction in serum albumin signal while still retaining hCG on the sampler during the washing protocol. An evaluation of the samplers revealed excellent linearity (R2 = 0.9995) for hCG in serum with relative standard deviations below 15%. This work has presented the first ever reported paper samplers immobilized with antibodies utilizing DVS chemistry, showing promise in the future of paper‐based sampling. [ABSTRACT FROM AUTHOR]

Background Monoclonal ( MoAb) reagents are routinely used and are usually very reliable for the serologic determination of ABO blood types. However, the fine specificity and cross-reactivity of these reagents are often unknown, particularly against synthetic antigens used in some diagnostic assays. If nonserologic assays or very sensitive techniques other than those specifically prescribed by the manufacturer are used, then there is a risk of incorrect interpretation of results. Study Design and Methods Forty-seven MoAbs and two polyclonal ABO reagents were tested against red blood cell ( RBC) kodecytes prepared with A trisaccharide, A Type 1, A Type 2, A Type 3, A Type 4, B trisaccharide, B Type 1, B Type 2, acquired B trisaccharide, and Lea trisaccharide function-spacer-lipid ( FSL) constructs. Natural RBCs were tested in parallel. In addition these FSL constructs were printed onto paper with a desktop inkjet printer and used in a novel immunoassay that identifies reactivity through the appearance of alphanumeric characters. Results Mapping of MoAbs with kodecytes and printed FSL constructs revealed a series of broad recognition patterns. All ABO MoAbs tested were reactive with the RBC dominant Type 2 ABO antigens. Unexpectedly some anti- A reagents were reactive against the B Type 1 antigen, while others were poorly reactive with trisaccharide antigens. Conclusions All ABO MoAbs detect the RBC dominant Type 2 ABO antigens; however, some reagents may show minor reactivity with inappropriate blood group antigens, which needs to be considered when using these reagents in alternative or highly sensitive analytic systems. [ABSTRACT FROM AUTHOR]

This perspective explores the feasibility of smart sampling with dried blood spots for the determination of proteins and peptides from human biomatrices using liquid chromatography coupled to mass spectrometry for clinical purposes. The focus is on innovative approaches to transform filter paper from a mere sample carrier to an active element in sample preparation, with the aim of reducing the need for extensive and intensive sample preparation in the conventional sense. Specifically, we discuss the use of modified cellulose to integrate sample preparation at an early stage of sample handling. The use of paper immobilized with either trypsin or monoclonal antibodies for protein digestion and affinity clean‐up is discussed as a potential benefit of starting sample preparation instantly at the moment of sampling to optimize time efficiency and enable faster analysis, diagnosis, and follow‐up of patients. [ABSTRACT FROM AUTHOR]

Whether the response to chemotherapy is a prognosticator in multiple myeloma (MM) is still not known. Therefore, the relationship between survival and the rate of monoclonal protein (M-protein) decrement during the first cycles of therapy was prospectively assessed in 262 patients with newly diagnosed MM that were included in a phase III trial (HOVON-16). M-proteins were collected monthly during melphalan–prednisone therapy (MP: melphalan 0·25 mg/kg, prednisone 1·0 mg/kg orally for 5 d every 4 weeks). Patients with light chain disease ( n = 18), immunoglobulin M (IgM)-MM ( n = 1) and no immunotyping ( n = 1) were excluded. Of the 242 patients studied, 75% had IgG M-protein and 25% IgA; MM stages: I: 1%, II: 35% and III: 64%. The median M-protein decrease after the first cycle of MP was 21% for IgG and 27% for IgA, and declined to < 5% after four cycles. An obvious survival advantage was seen for patients who had an M-protein decrease of at least 30% after the first MP cycle, which became significant when an M-protein decrease of 40% or more was reached. As established prognostic parameters (Salmon & Durie stage, serum creatinine, and haemoglobin) also remained prognostically significant, we concluded that early response to MP predicts for survival in MM. [ABSTRACT FROM AUTHOR]

Approximately 15% of patients undergoing non-myeloablative allogeneic haematopoietical cell transplantation (NMHCT) develop steroid-refractory acute-graft versus host disease (aGVHD), a usually fatal complication. We encountered 18 cases of steroid-refractory aGVHD in 146 patients, undergoing NMHCT from a related human leucocyte antigen-compatible donor following cyclophosphamide/fludarabine-based conditioning. Our initial cohort of steroid-refractory aGVHD patients treated with antithymocyte globulin (ATG) and mycophenolate mofetil (regimen-1: n = 6) had high GVHD-related mortality. Therefore, we investigated an alternative strategy for subsequent patients developing this complication (regimen-2: n = 12), consisting of daclizumab (alone or combined with infliximab/ATG) and targeted broad spectrum antibacterial and aspergillus prophylaxis in conjunction with rapid tapering of steroids to minimize opportunistic infections. In a retrospective analysis, patients receiving regimen-2 were significantly more likely to have complete resolution of GVHD compared with those receiving regimen-1 [12/12 (100%) vs. 1/6 (17%); P < 0·001]. When compared with those receiving regimen-1, regimen-2 patients also had a higher probability of survival at day 100 (100% vs. 50%) and day 200 (73% vs. 17%) post-transplant, and improved overall survival (median 453 d vs. 42 d from aGVHD onset; P < 0·0001). GVHD-related mortality was 89% for regimen-1 patients vs. 17% for regimen-2 patients ( P < 0·0001). These data suggest that a co-ordinated approach using immunoregulatory monoclonal antibodies, pre-emptive antimicrobial therapy and judicious steroid withdrawal can dramatically improve outcome in steroid-refractory aGVHD. [ABSTRACT FROM AUTHOR]

Novel antibodies have been generated by immunizing with bacterially expressed fragments of the repetitive motif of the Ki-67 gene. One such antibody, MIB1, recognizes a fixation and embedding resistant epitope on the Ki-67 protein if sections are previously microwaved in a citrate buffer. We have investigated the utility of this antibody as a marker of cell proliferation in archival material. The microwave technique is simple but requires careful monitoring since different tissues and fixatives require different irradiation times. Strong nuclear immunoreactivity was detected with all fixatives studied. Cytoplasmic staining was not identified. In a wide range of normal tissues the distribution and number of MIB1 immunoreactive cells matched that of cryostat sections stained with Ki-67. In nude mouse xenografts in which the growth fraction had been defined using a fraction of labelled mitosis method, the labelling index with MIB1 matched that previously determined for Ki-67 and correlated well with the growth fraction. Other markers of proliferation (e.g. proliferating cell nulcear antigen) have been shown to be expressed in DNA repair, thus we investigated expression of MIB1 immunoreactivity in situations of DNA repair in vivo—ultraviolet irradiated human skin. MIB1 staining correlated with semi-conservative DNA synthesis rather than excision repair DNA synthesis. Finally, the morphological and cell cycle distribution of MIB1 expression is identical to that of Ki-67. Thus, MIB1 represents a new anti-Ki-67 antibody which appears to be a robust marker of cell proliferation easily applicable to archival material. [ABSTRACT FROM AUTHOR]

Eighty cases of non-Hodgkin's lymphomas were examined independently using the monoclonal antibody Ki-67 and an argyrophilic method for the demonstration of nucleolar organizer regions. The evidence that Ki-67 immunoreactivity may be used as a marker of cell proliferation is described and the nature of nucleolar organizer regions reviewed, The proportion of tumour cells with nuclear Ki-67 immunoreactivity and the mean number of nuclear organizer regions are shown to be linearly related (r=0.86, P<0.001) although some scatter was observed. These data suggest that the mean number of nucleolar organizer regions may reflect the cellular kinetics of a tumour. This study also provides further evidence supporting the thesis that the mean nucleolar organizer region score is related to the histological grade of non-Hodgkin's lymphoma. Ki-67 immunostaining and nucleolar organizer region staining would seem to provide comparable data, at least in non-Hodgkin's lymphoma, but the latter method has the advantage of being applicable to conventionally fixed and processed paraffin sections. [ABSTRACT FROM AUTHOR]

Background: Lewis serologic reagents frequently give inaccurate phenotyping results. Furthermore these serologic reagents are often used in nonserologic assays such as inhibition and immunohistochemistry. In both scenarios knowledge of the fine specificity and cross-reactivity of these reagents will improve the quality of results obtained.Study Design and Methods: A range of contemporary and historical workshop and developmental Lewis reagents including mouse monoclonal (MoAb) and human and goat polyclonal (PoAb) reagents were evaluated. All were evaluated both against Lewis kodecytes expressing only single Le(a) , Le(b) , ALe(b) , BLe(b) , Le(x) , Le(y) , ALe(y) , or BLe(y) antigens and against the same antigens inkjet printed on a paper-based microplate and analyzed by enzyme immunoassay. Nine clinical samples were also evaluated. A kodecyte antigen dilution sensitivity assay was used to establish the ratio of Le(b) antigen between group A1 /A2 and O RBCs.Results: A continuum of cross-reactivity from Le(x) through to H was observed with MoAbs. All PoAb and few MoAb anti-Le(a) samples and reagents cross-reacted to some degree with Le(b) antigen. Some PoAb and MoAb anti-Le(b) did not cross-react with Le(a) . All polyclonal goat anti-Le(b) reagents showed substantial activity against ALe(b) and BLe(b) , while no MoAb reagent had this activity. A1 RBCs had less than half the Le(b) antigen of A2 /O RBCs.Conclusions: Substantial cross-reactivity of both MoAbs and PoAbs with related antigens highlights the risks of using serologic reagents in nonserologic assays or against synthetic antigens. The lack of ALe(b) activity in anti-Le(b) MoAbs explains their poor performance against blood group A1 Le(a-b+) phenotypes. [ABSTRACT FROM AUTHOR]

Over the past century, the field of antibody discovery has undergone significant evolution, excluding the current exploration stage of artificial intelligence‐based antibody generation and the often overlooked non‐animal sourced antibody discovery, which typically requires mature in vitro affinity and the selection of high‐quality antigen formulations. This journey has traversed various stages, from methods involving serum‐based antibody acquisition, the isolation of B cells capable of perpetual antibody production through hybridoma technology, to the in‐depth exploration of genetic material using the phage display system, and the current stage involving diverse single B cell screening techniques. Additionally, the emergence of machine learning has brought impressive scientific and technological breakthroughs across research domains, proving to be a powerful application in the field of antibody discovery. However, each technique comes with its limitations, such as variability and control challenges in serum‐based acquisition, lengthy and difficult hybridoma‐derived antibody development, potential limitations in sequence and epitope diversity due to immunization biases in phage display techniques, and costly single B cell screening. Protein mass spectrometry sequencing, with shorter acquisition time and lower costs, is seen as a shortcut by diagnostic companies, impacting traditional antibody development. In diagnostic antibody development, methodological differences in downstream assays and the impact of constant regions outside the Fv core are often neglected. This paper deeply analyzes challenges, proposing innovative strategies for the next generation of diagnostic antibody development. Aimed at moving closer to the gold standard of antibody discovery, these strategies enhance the competitiveness of diagnostic reagent products. [ABSTRACT FROM AUTHOR]

Cites several research papers on biochemistry. "Nematocysts (stinging capsules of Cnidaria) as Donnan-potential-dominated osmotic systems," by J. Weber; "Characterization of a high-affinity monoclonal antibody to calcineurin whose epitope defines a new structural domain of calcineurin A," by M.J. Hubbard and C.B. Klee; "Immunological detection of phytoene by bolulinum neurotoxin A - Contribution of various fragments to the intoxication process," by B. Poulain and colleagues.

Pharmacometric modelling plays a key role in both the design and analysis of regulatory trials in paediatric drug development. Studies in adults provide a rich source of data to inform the paediatric investigation plans, including knowledge on drug pharmacokinetics (PK), safety and efficacy. In children, drug disposition differs widely from birth to adolescence but extrapolating adult to paediatric PK, safety and efficacy either with pharmacometric or physiologically based approaches can help design or in some cases reduce the need for clinical studies. Aspects to consider when extrapolating PK include the maturation of drug metabolizing enzyme expression, glomerular filtration, drug excretory systems, and the expression and activity of specific transporters in conjunction with other drug properties such as fraction unbound. Knowledge of these can be used to develop extrapolation tools such as allometric scaling plus maturation functions or physiologically based PK. PK/pharmacodynamic approaches and well‐designed clinical trials in children are of key importance in paediatric drug development. In this white paper, state‐of‐the‐art of current methods used for paediatric extrapolation will be discussed. This paper is part of a conect4children implementation of innovative methodologies including pharmacometric and physiologically based PK modelling in clinical trial design/paediatric drug development through dissemination of expertise and expert advice. The suggestions arising from this white paper should define a minimum set of standards in paediatric modelling and contribute to the regulatory science. [ABSTRACT FROM AUTHOR]

Presents articles to be published in the "European Journal of Biochemistry." "An Antibody to the Low-Density Lipoprotein (LDL) Receptor That Partially Inhibits the Binding of LDL to Cultured Human Fibroblasts," by S. J. P. Gavigan, A. K. Soutar and B. L. Knight; "Association of Glyceraldehyde-3-Phosphate Dehydrogenase With Mono- and Polyribosomes of Rabbit Reticulocytes," by A. G. Ryazanov, L. I. Ashmarina and V. I. Muronetz; "Immunochemical Properties of Elongation Factors 1 of Plant Origin," by J. Pulikowska, P. Wojtaszek, A. Korcz, Z. Michalski, T. Candresse and T. Twardowski; "Use of a Monoclonal Antibody to Measure the Surface Expression of Thrombospondin Following Platelet Activation," by C. Legrand, V. Dubernard, N. Kieffer and A. T. Nurden; Others.

Risankizumab is a high‐affinity neutralizing anti‐interleukin (IL)‐23 monoclonal antibody marketed in over 40 countries across the globe to treat several inflammatory diseases, such as plaque psoriasis (PsO), psoriatic arthritis (PsA), and Crohn's disease (CD). This paper reviews the regulatory approval, mechanism of action, pharmacokinetics (PKs)/pharmacodynamics, immunogenicity, and clinical efficacy and safety data for risankizumab, focusing on the three main approved indications. Risankizumab binds to the p19 subunit of IL‐23 and inhibits IL‐23 from interacting with the IL‐23 receptor and subsequent signaling. Biomarker data obtained following treatment with risankizumab in multiple indications provided supportive evidence for downstream blockade of IL‐23 signaling associated with disease pathology. The PKs of risankizumab is linear and time‐independent, consistent with typical IgG1 monoclonal antibodies, across all evaluated indications. Risankizumab exhibited positive exposure‐response relationships for efficacy with no apparent exposure‐dependent worsening in safety. Immunogenicity to risankizumab had no major clinical consequences for either efficacy or safety. Efficacy and safety of risankizumab have been established in PsO, PsA, and CD in the pivotal clinical trials where superior benefit/risk profiles were demonstrated compared to placebo and/or active comparators. Moreover, safety evaluations in open‐label extension studies following long‐term treatment with risankizumab showed stable and favorable safety profiles consistent with shorter‐term studies. These data formed the foundation for risankizumab's marketing approvals to treat multiple inflammatory diseases across the globe. [ABSTRACT FROM AUTHOR]

This article focuses on a list of forthcoming papers published in the journal. Some of them are: "Conformation and Domain Structure of the Non-Histone Chromosomal Proteins," "Differential Response of Cultured Parsley Cells to Elicitors from Two Non-Pathogenic Strains of Fungi," "Monoclonal Antibodies Against Platelet Membrane Glycoproteins," "Synthesis of Biotin-labelled Dexamethasone Derivatives: Novel Harmone-Affinity Probes," "On the Mechanisms of ATP-induced and Succinate-Induced Redistribution of Cations in Isolated Rat Liver Cells," "Shedding of Gangliosides From Calf Thymocytes."

Presents several abstracts for articles presented during the Sixteenth Annual General Meeting of the Scandinavian Society for Immunology in Reykjavik, Iceland on June 12-16, 1985. "Monoclonal Antibodies Against Group O Streptococci," by H. Andersen, F. Larsen, C. Lindhardt, J. Selmer, and M. Wilken; "Low PPD-Induced Lymphocyte Stimulation Response in HTLV-III/LAV Infection is Related to Monocytes, But Not to Lymphocytes," by Jaakko Antonen and Kai Krohn.

The epithelial cell adhesion molecule (Ep-CAM) is expressed on the surface of most human carcinomas, including ovarian, breast, lung, prostate and colorectal carcinoma. Ep-CAM was shown to be a valid target for monoclonal antibody-based therapies. We have investigated whether an Ep-CAM-/CD3-bispecific single-chain antibody called bscEp-CAM × CD3 is effective in tumor cell elimination within the cellular microenvironment of primary ovarian cancer tissue. The ex vivo elimination of ovarian cancer cells in tumor preparations from 21 patients was monitored by flow cytometry using Ep-CAM/CA-125 double-labeling or Ep-CAM single-labeling combined with propidium iodide uptake of cells. Methodology was established by the ovarian cancer cell line OvCAR. A total of 17 (81%) patient samples showed a dose-dependent tumor cell elimination by bscEp-CAM × CD3. High and specific tumor cell lysis was seen at bscEp-CAM × CD3 concentrations as low as 1 ng/ml, at very low effector:target ratios and in the absence of T cell costimulation. The high efficacy of the bispecific antibody may be due to the non-restricted activation of tumor-resident cytotoxic T lymphocytes. In clinical trials, the ex vivo data with the T cell-recruiting bispecific antibody bscEp-CAM × CD3 may translate into a high response rate and efficacy of tumor cell elimination. © 2003 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Certain therapies that target CD markers on some blood cells can affect pretransfusion testing. Key examples are anti‐CD38, CD47 monoclonal antibody (mAb) therapies such as daratumumab (DARA) and magrolimab, which have presented a challenge for transfusion medicine laboratories around the globe. Scientists have been faced with not only introducing a protocol to provide safe blood to patients but also investigating the most effective method to remove the pretransfusion pan‐agglutinating interference caused. A number of papers in the last 5 years have reported on various methods to remove pretransfusion interference; however, most of these studies have been conducted only in a few countries. Most recent reviews on this topic have focused on techniques and reagents to remove pretransfusion interference, and dithiothreitol is currently the gold standard for removing DARA interference. However, it was clear from this review that while many laboratories have developed processes for addressing interference in pretransfusion testing, and DARA interference may not be a major issue, there are still laboratories around the world, that may not have adequately addressed this issue. In addition, the impact of mAb interference on widely used techniques such as flow cytometry is unclear. [ABSTRACT FROM AUTHOR]

Membrane fouling by monoclonal antibodies (mAbs) is one of the main challenges in virus‐filtration processes. Previous publications attributed membrane fouling to the presence of mAb aggregates in the solution, which block the membrane pores. This fouling mechanism can be solved by a prefilter; however, it was shown that there are mAbs that severely foul the membranes (reduce permeability by 90% and more) even after prefiltering the aggregates, while other mAbs foul the membrane weakly (reduce permeability by ~10% and less). Unfortunately, the differences between the fouling‐ and the nonfouling mAbs have never been convincingly explained. To get a deeper insight on these differences, we measured the fouling of chemically modified Isoprene‐Styrene‐4‐vinylpyridine (ISV) membranes (TeraPore Technologies) by 8 mAbs exhibiting different hydrophobicity and charge. The results show that mAb solutions with low concentration of aggregates foul ISV membranes via an adsorptive mechanism, and the adsorption is driven mainly by hydrophobic forces between the mAb and the membrane. The charge of the mAbs plays a secondary role in fouling. We want to emphasize that the conclusions pertain to ISV membranes; the insights presented in this paper can potentially be used to engineer new surface chemistries to mitigate fouling of other virus‐filtration and/or ultrafiltration membranes. [ABSTRACT FROM AUTHOR]

In solid organ transplantation (SOT), biologicals such as recombinant therapeutic proteins, monoclonal antibodies, fusion proteins and conjugates are increasingly used for immunosuppression, desensitization, ABO (blood group) incompatibility, antibody‐mediated rejections and atypical haemolytic uremic syndrome. In this paper, we review the medical evidence available for biologicals used in SOT and the potential for improvement by the application of therapeutic drug monitoring (TDM) and model‐informed precision dosing. Biologicals are used for off‐label indications within the field of SOT, building on the experience from their use on labelled indications. Dosing is currently mostly standard, and experience vs. effect and toxicity is limited. Pharmacokinetic characteristics of these large, partly also immunogenic molecules differ from those of traditional small molecules. Individualization by concentration measurements and modelling has mostly been proof‐of‐concept or feasibility studies that lack the power to provide evidence for improvement in clinical outcome. For some drugs such as alemtuzumab, eculizumab, rituximab, tocilizumab and belatacept, studies have demonstrated significant interindividual variability in pharmacokinetics. Variability in absorption from subcutaneous administration may increase interindividual variability. There is also an economic aspect of appropriate dosing that needs to be pursued. Available assays and models to refine interpretation are in place, but trials of adequate size to document the usefulness of TDM and MIPD are scarce. Collaboration within the TDM community seems mandatory to establish studies of sufficient strength to provide evidence for the use of biologicals that are currently used off‐label in SOT and furthermore to identify the settings where TDM may be beneficial. [ABSTRACT FROM AUTHOR]

This paper presents an approach based on triple injection capillary zone electrophoresis for identification of monoclonal antibodies. The analyte to be identified is injected between two zones of a known reference. The distances between the reference zones (plug I and III) and the target zone (plug II) are adjusted by partial electrophoresis of the first and second injection plugs. The full migration time of the target analyte is calculated from the observed migration time by considering the migration times of the reference in the first and third injection plugs. The relative migration time, that is, the ratio between the full migration time of the analyte and the migration time of the reference in the third injection plug provides the basis for identification. Here, eight monoclonal antibodies, including a pair of biosimilars, were used interchangeably as both analyte and reference to investigate potential of the method. The relative migration time for a preliminary positive identification were found to vary between 0.994 and 1.006 (1.000 ± 0.006, p = 95%). Beside the relative migration time, isoform distribution, peak profiles, and early migrating peaks, originating from components in the pharmaceutical formulations, were successfully used to verify the identity of all tested monoclonal antibodies. [ABSTRACT FROM AUTHOR]

Light chain amyloidosis is a rare disease caused by clonal plasma cells in the bone marrow generating an excessive amount of immunoglobulin light chains. These chains misfold and produce insoluble fibrils that deposit in various organs, including the heart, kidneys, liver, nervous system, and digestive tract. Life expectancy and symptoms during the course of the disease vary depending on which and how many organs are affected. Targeted plasma cell therapy has significantly advanced the clinical management of amyloidosis, with ongoing progress. However, current clinical studies are investigating innovative targets, drug combinations and treatment strategies to improve therapeutic outcomes by minimizing adverse effects and refining patient prognosis in these challenging hematological conditions. In this paper, we review the state of the art regarding the use of anti‐amyloid antibodies, as a revolutionary and innovative approach in the current scenario of amyloid treatment. [ABSTRACT FROM AUTHOR]

Teaching chemistry and biology students about biologics design remains challenging despite its increasing importance in pharmaceutical development. Monoclonal antibodies, commonly called mAbs, are the most popular biologics. They have been developed into drugs to treat various diseases in the past decades. Multiple challenges exist for designing proper formulations to stabilize mAbs, such as preventing aggregation and mitigating viscosity. Molecular modeling and simulations can improve pharmaceutical products by examining the interactions between mAbs and other compounds, such as excipients. To introduce students to biopharmaceuticals, eight students at the Stevens Institute of Technology participated in a semester‐long course to learn the challenges of pharmaceutical development and different computational skills to study biologics design. The students started with a limited background in this field. Throughout one semester, they were introduced to various literature and software tools for modeling antibodies and studying their interactions with excipients. This paper aims to develop a course structure to be replicated at other universities and institutions to teach biopharmaceutical development to students. [ABSTRACT FROM AUTHOR]

Objective: To compare the effectiveness and safety of galcanezumab, fremanezumab, and erenumab for the treatment of chronic and episodic migraine, through real‐world data. Background: Monoclonal antibodies (mAbs) targeting the calcitonin gene–related peptide (CGRP) pathway have been tested extensively in several clinical trials for both episodic and chronic migraine, showing high effectiveness, safety, and tolerability; however, there are no prospective real‐world studies intending to compare their efficacy and safety. Methods: This is a prospective observational cohort study comparing the effectiveness and safety profiles of galcanezumab, fremanezumab, and erenumab for the treatment of chronic and episodic migraine. We enrolled 140 patients at the Headache Centre of University Federico II of Naples, with a history of multiple failed treatments with validated migraine preventatives. Framenezumab, erenumab, or galcanezumab were administered for 12 months. The mean monthly days with headache, Migraine Disability Assessment (MIDAS) score, and adverse events were evaluated during the run‐in period and every 3 months by reviewing standardized paper patient headache diaries. Results: We found a mean reduction of migraine monthly days from baseline of −12.0 (−9.8, −14.1) in the galcanezumab group, −12.3 (−10.2, −14.3) in the fremanezumab group, and −10.8 (−8.5, −13.1) in the erenumab group (for all, p < 0.001). We found a mean reduction of MIDAS score of −32.6 (−26.6, −38.5) in the galcanezumab group, −33.4 (−28.0, −38.9) in the fremanezumab group, and −29.2 (−23.0, −35.4) in the erenumab group (for all, p < 0.001). We found no significant differences between mAbs in the reduction of mean monthly days with headache and MIDAS score. We found a more rapid effect of galcanezumab and erenumab compared to fremanezumab in medication overuse headache patients after 3 months of treatment (−10.8 and −11.1 vs. −4.0 days; p = 0.029). Conclusion: Our results confirm the therapeutic benefits of anti‐CGRP mAbs. There is no evidence that suggests that one antibody may be superior to the others in terms of effectiveness, both in chronic and episodic patients. [ABSTRACT FROM AUTHOR]

Biological therapeutics are increasingly being formulated to high protein concentrations to decrease drug substance storage space and increase the flexibility of administration to patients. With the higher protein concentration targets comes added challenges to the downstream purification manufacturing process. Tangential flow filtration (TFF) operations are typically performed to reach target protein concentration. TFF operations for a given drug substance may include an Ultrafiltration and Diafiltration (UFDF) step, or a UFDF step followed by a single pass tangential flow filtration (SPTFF) step. Whether a TFF step achieves a target protein concentration is determined by at‐line protein concentration measurements performed at the completion of a process step. If the measured protein concentration is outside the specified range, the unit operation may need to be restarted, reprocessing may need to be performed, or a batch may need to be terminated. Out of specification protein concentration measurements may be a result of the TFF operation or sample measurement. Increased viscosities associated with high concentration TFF operations pose added challenges to the TFF process and sample measurement. Implementation of an inline process analytical technology (PAT) to monitor real‐time protein concentration during TFF operations has the potential to improve the accuracy of the operations in achieving target protein concentrations. This will result in improved process consistency and efficiency, increased operator confidence and decreased likelihood of batch failures. This paper studies the performance of a K‐Patents PR‐23 refractometer (Vaisala) as a PAT to monitor and control the UFDF and SPTFF unit operations of a commercial scale monoclonal antibody purification process. [ABSTRACT FROM AUTHOR]

Sakiyama Y, Kato R, Inoue S, Suzuki K, Itabe H, Yamamoto M. Detection of oxidized low-density lipoproteins in gingival crevicular fluid from dental patients. J Periodont Res 2009; doi: 10.1111/j.1600-0765.2009.01226.x. © 2009 John Wiley & Sons A/S Background and Objective: Oxidative modification of low-density lipoprotein (LDL) occurs in various diseased tissues and sites of local inflammation. For example, an increased plasma oxidized low-density lipoprotein (OxLDL) level is a well-known risk marker for cardiovascular diseases. Gingival crevicular fluid, the exudate from gingival tissues into the sulci, can be easily collected in a non-invasive manner. However, the possible presence of OxLDL in gingival crevicular fluid has not been studied. In this study, we established a procedure to measure OxLDL in human gingival crevicular fluid. Material and Methods: Human gingival crevicular fluid was sampled with paper points or paper strips. The gingival crevicular fluid samples from healthy gingival sulci (pocket depth < 4 mm, n = 14) were subjected to western blot and/or sandwich ELISA. The amounts of OxLDL and LDL were measured by sandwich ELISA using an anti-oxidized phosphatidylcholine monoclonal antibody and two anti-apolipoprotein B antibodies. Venous blood samples were analyzed biochemically. Results:  We tested two methods of gingival crevicular fluid collection, namely paper points and paper strips. Gingival crevicular fluid could be collected very safely with paper points and they showed good recovery of LDL and OxLDL throughout the analysis. Apolipoprotein B, the major protein component in LDL, was detected in gingival crevicular fluid by western blot, and OxLDL was found to be present in gingival crevicular fluid by ELISA. The OxLDL/LDL ratio in gingival crevicular fluid was 17.0 times higher than that in plasma. Conclusion: This is the first report to show the presence of apolipoprotein B and apolipoprotein B– oxidized phosphatidylcholine complex, which correspond to LDL and OxLDL, respectively, in gingival crevicular fluid. [ABSTRACT FROM AUTHOR]

Leo Augustus Behie was known as a dedicated teacher and mentor, a team builder, and an outstanding researcher and innovator. He had a most productive and illustrious career, spanning five decades, that covered both industrial and academic sectors. His academic research career started with tackling classical chemical engineering challenges and, with time, shifted to biotechnology and biomedical engineering. He was the founding director of a unique research laboratory for undertaking innovative biotechnology and biomedical engineering research at the University of Calgary, called the Pharmaceutical Production Research Facility (PPRF). This paper presents an overview of his academic and research accomplishments, which also included the training of 17 doctoral students, 21 master's degree students, and many postdoctoral scholars. His main research areas under the classical chemical engineering theme included the investigation of the grid region in gas–solid fluidized bed reactors; gas–liquid and liquid–solid multiphase systems; fluidized bed reactors for metals recovery from coal ash; spouted fluid‐bed reactors; kinetics of pyrolysis chemical reactions; transport phenomena in rotary drums; and reactions, thermodynamics, and reactor modelling of sulphur systems. His main research areas under the biotechnology and biomedical engineering theme included advances in biotechnology and bioreactor design; production of antibiotic and monoclonal antibody; protein production in insect cells; process development for the production of neural stem cells, cancer stem cells, and islet cells; mesenchymal stem cells; and cellular secretome studies aimed at developing a cure for Parkinson's disease. [ABSTRACT FROM AUTHOR]

In recent years, mRNA‐based therapeutics have been a fast‐growing new class of biologics that can, in principle, encode any protein(s) directly in patients to treat various diseases. mRNA vaccines have been proven to work efficiently, have high potency, and can be rapidly developed and deployed, which is critical for a quick responses in the case of a pandemic. Such agile development is enabled by rapid synthesis of RNA in vitro using recombinant enzymes rather than relying on lengthy and complex cell culture processes. mRNA exhibits physical and chemical properties differing from protein‐based therapeutics. It is highly negatively charged and the hydroxyl group makes mRNA less stable and more susceptible to hydrolysis and nucleophilic cleavage. This novel work shares comprehensive studies carried out to compare the performance of various mRNA purification strategies by considering its scalability and critical quality attributes. In addition, the paper provides insights on how to establish a scalable mRNA purification process that consists of ultrafiltration/diafiltration and chromatography steps with good recoveries. Alternative Oligo(dT) based columns were further explored aiming to improve total process recovery. With Oligo(dT) as a capture step, overall recoveries of 70% can be achieved for mRNAs studied here that encode anti‐influenza immunoglobulin G monoclonal antibodies. [ABSTRACT FROM AUTHOR]

With the approval of the first vaccines against respiratory syncytial virus (RSV) and a novel RSV‐neutralizing antibody, 2023 has been perceived as a game‐changing year in preventing severe outcomes of RSV infections in infants and the elderly. However, the costs of these pharmaceuticals are high, while RSV disproportionately impacts populations of low‐to‐middle‐income regions, which may continue to suffer from a lack of pharmaceutical measures for RSV prevention under health and socioeconomic disparities. This paper presents an overview of the characteristics, clinical results, and approval status of various RSV vaccines and anti‐RSV antibodies. It posits that wealthy nations cannot monopolize RSV immunoprophylaxis and should work jointly to make it available to lower‐income countries. An approach toward RSV immunoprophylaxis equity based on five points is offered: (1) integration of RSV vaccines and antibodies into the existing global humanitarian distribution systems, (2) using affordable RSV vaccine pricing models, (3) enforcing equity as a part of national and global public health strategy, (4) implementing equitable allocation frameworks for RSV immunoprophylaxis, and (5) promoting local manufacturing. Such a plan needs to be put into action as soon as possible to avoid delays in serving the populations with the highest needs related to RSV burden. [ABSTRACT FROM AUTHOR]

Hydrophobic interaction chromatography (HIC) is a chromatographic technique that mainly targets the separation of biomolecules (intact proteins, monoclonal antibodies, etc.) based on the difference in surface hydrophobicity while applying non‐denaturing conditions. This protocol paper provides guidelines for setting‐up robust HIC analysis and considers the instrument configuration, mobile‐phase and sample preparation, as well as chromatographic conditions and settings. The separation of a mixture of intact proteins and monoclonal antibodies is demonstrated by applying conventional HIC conditions, that is, using a mildly hydrophobic (C4) stationary phase in combination with an inverse ammonium sulphate gradient dissolved in aqueous phosphate buffer. The effect of sample‐preparation conditions on sample breakthroughs is presented. Finally, good run‐to‐run repeatability (relative standard deviation < 2%) is demonstrated for five different columns obtained from three different column lots, considering chromatographic retention, peak width, peak area and column pressure. [ABSTRACT FROM AUTHOR]

Objectives: This paper reviews current and emerging therapies for multiple myeloma (MM). Methods: Narrative review. Results: MM is a complex, heterogenous condition, and in recent years there has been an expansion in the number and range of treatments. Several new treatment approaches, including enhanced monoclonal antibodies, antibody‐drug conjugates, bispecific T‐cell engagers, and chimeric antigen‐T‐cell therapy are under development. Conclusions: The emergence of new treatments that aim to tackle MM‐associated immune dysfunction has led to improvements in overall survival. [ABSTRACT FROM AUTHOR]

LINKED CONTENT: This article is linked to D'Haens et al paper. To view this article, visit https://doi.org/10.1111/apt.17557 [ABSTRACT FROM AUTHOR]

Objective: To determine the effectiveness and safety of erenumab in patients with chronic migraine in the real‐world setting of 3 headache centers in Australia. Methods: Patients with migraine were prescribed erenumab (70 or 140 mg) in the setting of either a product familiarization program or paid access to the medication in 3 headache centers in Australia. We obtained baseline and monthly prospective data on monthly headache days, monthly migraine days, monthly triptan use days, monthly codeine use days, Headache Impact Test‐6 scores, and adverse reactions. In this paper, we present our data at 3 and 6 months in our subgroup of patients with chronic migraine with and without medication overuse. Results: A total of 170 patients with chronic migraine were prescribed erenumab in the 3 headache centers. At 3 months, 100/170 (58.8%) had 50% or greater reduction in monthly migraine days. At 6 months, 79/170 (46.5%) had 50% or greater reduction in monthly migraine days. At 6 months, there was a mean reduction in monthly headache days of 9.2 days, a mean reduction in monthly migraine days of 10.2 days. There were few adverse events reported. Conclusion: This is the first report from 3 Australian headache centers about erenumab in the real world. Our analysis has supported erenumab as an effective and well‐tolerated migraine preventative therapy for patients with chronic migraine who have failed many preventative therapies. [ABSTRACT FROM AUTHOR]

BACKGROUND Monoclonal antibodies (mAbs) presently dominate the biotherapeutic market. However, biotherapeutics in general, and mAbs in particular, are expensive therapies and outside the realm of affordability for most of the world. In an attempt to lower their cost of manufacturing, a novel in‐house non‐protein A continuous platform has recently been reported for mAb purification. In this paper, we perform an in‐depth evaluation of the process economics to assess the impact of removing the protein A step on the cost of goods (COGs). RESULTS: The non‐protein A platform was found to result in significant savings in COGs at about 23% when comparing both platforms in batch mode. Conversion of the non‐protein A platform from batch to continuous mode led to a further increase in these savings with about 39% gains in COGs over the conventional protein A platform. CONCLUSIONS: The non‐protein A purification platform offers a viable alternative to the standard protein A platform and can be considered when setting up a new facility. The continuous non‐protein A platform can especially be considered given the flexibility of the setup and the utility at both small and large scales. © 2021 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Objectives: The treatment goal for rheumatoid arthritis has undergone a dramatic change over recent years with the advent of biologic and targeted synthetic disease‐modifying antirheumatic drugs (b/tsDMARDs). This systematic review aims to discuss the comparative efficiency of new drugs versus the standard of care methotrexate. Methods: Pubmed, Google Scholar, and Science Direct were searched, and 7072 records were retrieved. Randomized controlled trials published in the last five years with full‐text papers, blinded studies, and studies with an open‐label extension were included. Studies with a high risk of bias as calculated by the Cochrane risk of bias (RoB2) tool were excluded. The last database search was conducted on 21 August 2021. Results: Fifteen studies were identified and analyzed by categorizing drugs into Janus kinase (JAK) inhibitors, tumour necrosis factor (TNF) inhibitors, and monoclonal antibodies with other targets (anti‐GM‐CSF receptor, anti‐IL6 receptor, anti‐IL‐17 monoclonal antibodies), with a total of 7963 participants. Data were synthesized with tables and the PRISMA flow diagram. Our analysis included baricitinib, filgotinib, peficitinib, infliximab, certolizumab, sarilumab, tocilizumab namilumab, mavrilimumab, golimumab, and CNT06785. Conclusions: These studies showed that except for CNT 06785 (anti‐Interleukin 17A monoclonal antibody), biologic and targeted synthetic disease‐modifying drugs combined with methotrexate or as monotherapy yield better clinical and radiographical outcomes. This systematic review does not discuss biosimilars and the adverse effects of biologic drugs in detail. Infections remain a significant adverse drug reaction of monoclonal antibodies. However, these drugs play a crucial role in adequately managing this disease for patients with severe arthritis. [ABSTRACT FROM AUTHOR]

High‐throughput screening (HTS) approaches are commonly used to accelerate downstream process development. Although most HTS approaches use batch isothermal data (KP screen) or bind and elute mode as screening procedure, different or new process designs are rarely investigated. In this paper, a mechanistic model case study for the separation of two different two‐component solutions was conducted and confirmed prior evidence. With these outcomes, a novel HTS screening procedure was developed including the determination of competitive adsorption‐based displacement effects and key parameter identification. The screening procedure employing an overload bind and elute (OBE) mode is presented in a case study dealing with IgG aggregate removal in a typical monoclonal antibody purification step, applying a Sartobind® S membrane adsorber (MA). Based on a MA scale down device, the OBE mode allows the determination of classical process parameters and dynamic effects, such as displacement effects. Competitive adsorption‐based displacement effects are visualized by introducing a displacement identifier leading to a displacement process map. Based on this map, the approach is transferred to and confirmed by the OBE recycle experiments with 4.6 and 8.2 ml benchtop scsale devices resulting in 45% reduced IgG monomer and 88% increased higher molecular weight species binding capacities. [ABSTRACT FROM AUTHOR]

Monoclonal antibodies and antibody fragments are increasingly important classes of biotherapeutics. However, these products are both challenging and expensive to manufacture. New process analytical technologies used to monitor these products during their manufacture are of significant interest. Deep UV Raman spectroscopy promises to provide the required specificity and accuracy, however instruments, have historically been large and complex. In this paper, a new deep UV Raman instrument is described using a solid‐state laser and a spatial heterodyne spectrometer. The instrument overcomes practical limitations of the technique and could readily be used for online measurement. A series of observations have been made of biopharmaceutical products, including immunoglobulin G and domain antibodies. Where high levels of both specificity and linearity when measuring samples of different concentration with a precision of better than 0.05 mg/mL has been demonstrated. [ABSTRACT FROM AUTHOR]

An increased level of granulocyte‐macrophage colony‐stimulating factor has a potential role in the development of autoimmune diseases, and the neutralization of its activity by monoclonal antibodies is a promising therapy for some diseases. Here, the crystal structure of the Fab region of EV1007, a fully human antibody expressed in Chinese hamster ovary cells that was developed from human peripheral blood mononuclear cells, is described. The structure closely resembles that of MB007, which is the Fab region of the same antibody expressed in Escherichia coli [Blech et al. (2012), Biochem. J.447, 205–215], except at the hinge regions between the immunoglobulin domains and the H3 loop region. This paper presents evidence for the flexibility of the hinge and H3 loop regions of the antibody based on the comparison of two independently solved crystal structures. [ABSTRACT FROM AUTHOR]

Recently, there have been many studies and reports on various continuous protein crystallization processes. Due to the increasing demand for pharmaceutical proteins such as insulin and monoclonal antibodies, it has become essential to optimize the current industrial processes to meet these demands. There are various comparisons to be made between current batch crystallization processes and continuous processes for proteins. This review paper aims to review the recent advancements in continuous protein crystallization processes, such as slug flow crystallizers, oscillatory baffled crystallizers, and mixed suspension mixed product removal (MSMPR) crystallizers. [ABSTRACT FROM AUTHOR]

This paper discusses ultrahigh‐speed, ultrahigh‐resolution preparative protein separation using an in‐house designed membrane chromatography device. The performance of the membrane chromatography device was systematically compared with an equivalent resin‐packed preparative column. Experiments carried out using model proteins showed that membrane chromatography gave more than four times greater resolution than the preparative column, while at the same time being more than 19 times faster. Membrane chromatography was therefore a better option, not only in terms of higher productivity but also in terms of higher product purity. Membrane chromatography was also superior in terms of resolving and presenting tracer impurity peaks in the chromatogram. Experiments carried out using monoclonal antibody samples showed that membrane chromatography was suitable for ultrahigh speed, and ultrahigh resolution fractionation of charge variants. This paper highlights and explains the need for proper device design for enabling the use of membrane chromatography for the efficient purification of protein biopharmaceuticals. [ABSTRACT FROM AUTHOR]

Cellular therapy nowadays includes various products from haematopoietic stem cells (HSC) collected from bone marrow, peripheral blood, and umbilical cord blood to more complex adoptive immune therapy for the treatment of malignant diseases, and gene therapy for inherited immune deficiencies. Broader utilization of cellular therapy requires extensive quality testing of these products that should fulfil the same requirements regarding composition, purity, and potency nevertheless they are manufactured in various centres. Technical improvements of the flow cytometers accompanied by the increased number of available reagents and fluorochromes used to conjugate monoclonal antibodies, enable detailed and precise insight into the function of the immune system and other areas of cell biology, and allows cell evaluation based on size, shape, and morphology or assessment of cell surface markers, as well as cell purity and viability, which greatly contributes to the development and progress of the cell therapy. The aim of this paper is to give an overview of the current use and challenges of flow cytometry analysis in quality assessment of cellular therapy products, with regard to basic principles of determining HSC and leukocyte subpopulation, assessment of cells viability and quality of thawed cryopreserved HSC as well as the importance of validation and quality control of flow cytometry methods according to good laboratory practice. [ABSTRACT FROM AUTHOR]

Summary: Early detection of expanded‐spectrum cephalosporin (ESC) resistance is essential not only for an effective therapy but also for the prompt implementation of infection control measures to prevent dissemination in the hospital. We have developed and validated a lateral flow immunoassay (LFIA), called LFIA‐CTX test, for the detection of ESC (cefotaxime) hydrolytic activity based on structural discrimination between the intact antibiotic and its hydrolysed product. A single bacterial colony was suspended in an extraction buffer containing cefotaxime. After a 30‐min incubation, the solution is loaded on the LFIA for reading within 10 min. A total of 348 well‐characterized Gram‐negative isolates were tested. Among them, the 38 isolates that did not express any cefotaxime‐hydrolysing β‐lactamase gave negative results. Of the 310 isolates expressing at least one cefotaxime‐hydrolysing β‐lactamase, all were tested positive, except three OXA‐48‐like producers, which were repeatedly detected negative. Therefore, the sensitivity was 99.1% and the specificity was 100%. The LFIA‐CTX test is efficient, fast, low‐cost and easy to implement in the workflow of a routine microbiology laboratory. This paper details the development and validation of a lateral flow immunochromatographic assay for the detection of hydrolysis activity of extended spectrum cephalosporins. This test, based on the use of monoclonal antibodies specific to intact antibiotics, takes advantage of the LFIA and the universality of biochemical tests (detection of enzymatic activity whatever the enzyme or its variants). Its validation on 348 bacterial strains indicates a sensitivity of 99.1% and a specificity of 100%, making it an efficient, rapid, low‐cost and easy to implement test in the workflow of a routine microbiology laboratory, without additional equipment or trained personnel. [ABSTRACT FROM AUTHOR]

OX40 is a costimulatory molecule that belongs to the tumor necrosis factor receptor (TNFR) superfamily. OX40 agonist‐based combinations are emerging as promising candidates for novel cancer immunotherapy. Clinical trials have shown that OX40 agonist antibodies could lead to better results in cancer patients. Using a hybridoma platform and three different types of immunization strategies, namely recombinant protein, DNA, and overexpressing cells, we identified a chimeric anti‐OX40 antibody (mAb035‐hIgG1 from DNA immunization) that shows excellent binding specificity, and slightly stronger activation of human memory CD4+ T cells and similar potent antitumor activity compared with BMS 986178, an anti‐OX40 antibody currently being evaluated for the treatment of solid tumors. This paper further systematically investigates the antigen‐specific immune response, the number of binders, epitope bins, and functional activities of antibodies among different immunization strategies. Interestingly, we found that different immunization strategies affect the biological activity of monoclonal antibodies. [ABSTRACT FROM AUTHOR]

Viral infections cause high morbidity and mortality, threaten public health, and impose a socioeconomic burden. We have seen the recent emergence of SARS‐CoV‐2 (Severe Acute Respiratory Syndrome Coronavirus 2), the causative agent of COVID‐19 that has already infected more than 29 million people, with more than 900 000 deaths since its identification in December 2019. Considering the significant impact of viral infections, research and development of new antivirals and control strategies are essential. In this paper, we summarize 96 antivirals approved by the Food and Drug Administration between 1987 and 2019. Of these, 49 (51%) are used in treatments against human immunodeficiency virus (HIV), four against human papillomavirus, six against cytomegalovirus, eight against hepatitis B virus, five against influenza, six against herpes simplex virus, 17 against hepatitis C virus and one against respiratory syncytial virus. This review also describes future perspectives for new antiviral therapies such as nanotechnologies, monoclonal antibodies and the CRISPR‐Cas system. These strategies are suggested as inhibitors of viral replication by various means, such as direct binding to the viral particle, blocking the infection, changes in intracellular mechanisms or viral genes, preventing replication and virion formation. We also observed that a large number of viral agents have no therapy available and the majority of those approved in the last 32 years are restricted to some groups, especially anti‐HIV. Additionally, the emergence of new viruses and strains resistant to available antivirals has necessitated the formulation of new antivirals. [ABSTRACT FROM AUTHOR]

The amino acid sequence of the light chain of a human monoclonal IgA1 (Mem) was established, in part by analogy with already known sequences. By homology its variable part was shown to belong to the VλI subgroup while the isotype-associated amino acid residues characterized it as Mcg+, Kern+ and Oz-. The normal primary structure of this chain was in contrast to its abnormal physical and antigenic properties: (a) its apparent molecular mass estimated by SDS/polyacrylamide gel electrophoresis, by gel filtration chromatography and by gradient ultracentrifugation was found to be lower by &assymp; 10% than the values (23.5 kDa) of ‘normal’ light chain used as controls; (b) the λI chain Mem, when tested in native state was not antigenically reactive. These abnormalities were reverted when the chain was treated with 8 M urea. These data suggest that the abnormal behaviour of λI chain Mere is at a conformational level. [ABSTRACT FROM AUTHOR]

Background: Asthma is one of the most common chronic diseases worldwide and can often be controlled by existing treatments, but in the 5%‐10% of patients with severe asthma, control is frequently more challenging. The aim of this paper is to evaluate the adherence to preventer inhaler therapy in severe asthmatic patients receiving omalizumab, a recombinant humanized monoclonal anti‐IgE antibody indicated for moderate‐to‐severe asthma. Methods: This study included 29 patients who were receiving omalizumab treatment as maintenance therapy. All patients completed the six question Turkish Modified Morisky Scale. In addition, patients were classified by adherence to their prescribed preventer therapy, based on pharmaceutical and medical device institution records from the preceding one year. Results: Of the 29 enrolled patients, 24.1% had NSAID‐exacerbated respiratory disease; 75.9% were female and median age was 47.2 ± 12.99 years. Severe asthma patients demonstrated high levels of motivation (98.8%, n = 28) and knowledge (86.2%, n = 22) about asthma treatment. Nevertheless, the rate of nonadherence to preventer inhaler therapy was only 34.5% (n = 10). There were no significant differences among the characteristics of the patients that might affect the adherence to inhaler therapy. Conclusion: In patients with severe asthma, suboptimal adherence to preventer inhaler therapy is common before and after omalizumab treatment. Although patients have high levels of motivation and knowledge about asthma treatment, adherence to inhaler preventer therapy declined after omalizumab therapy. Given these findings, it is important to decide whether preventer inhaler therapy should be continued as monotherapy or be discontinued inpatients exhibiting impaired adherence to biological agents, such as omalizumab. [ABSTRACT FROM AUTHOR]

BACKGROUND: The lifetime of chromatography resins typically averages between 10 and 300 cycles for the manufacture of a therapeutic protein. Developing and establishing the robustness of the method for each separation process represents a significant challenge and is subject to extensive regulatory oversight. This paper presents a novel fluorescence‐based assay for residual aggregated proteins to aid the evaluation of the extent of resin regeneration. RESULTS: The versatility of this method was demonstrated by using strong anion and cation exchange agarose resins Praesto Q and SP in conjunction with bovine serum albumin and monoclonal antibody feed materials. The assay entails applying a molecular rotor dye to a sample of free resin and measuring the fluorescence intensity using a plate reader or visualizing under a confocal laser scanning microscope to gain a more detailed characterization. Following five consecutive chromatography cycles, both methods revealed a 10‐fold increase in fluorescence intensity along with a proportional reduction in dynamic binding capacity. Furthermore, the use of the assay suggested that fouling was dependent on spatial bead position in the column, bead channel structure and cleaning conditions. CONCLUSION: This work presents a simple assay suitable for use in resin lifetime studies to enhance process understanding. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Keywords: mass spectrometry; MGUS; plasma cell disorders; screening EN mass spectrometry MGUS plasma cell disorders screening 722 724 3 08/18/23 20230815 NES 230815 The emergence of screening for MGUS and mass spectrometry-based diagnostics could lead to a fundamental shift in the field of plasma cell disorders. We are currently conducting the first randomized study evaluating the benefits and harms of MGUS screening, the Iceland screens, treats or prevents MM study (iStopMM).[9] iStopMM is a nationwide population-based screening study for MGUS and subsequent randomized controlled trial of follow-up strategies. Mass spectrometry for the evaluation of monoclonal proteins in multiple myeloma and related disorders: an international Myeloma Working Group Mass Spectrometry Committee Report. [Extracted from the article]

Broadly neutralizing antibodies are proposed as therapeutic and prophylactic agents against HIV‐1, but their potency and breadth are less than optimal. This study describes the immunization of a llama with the prefusion‐stabilized HIV‐1 envelope (Env) trimer, BG505 DS‐SOSIP, and the identification and improvement of potent neutralizing nanobodies recognizing the CD4‐binding site (CD4bs) of vulnerability. Two of the vaccine‐elicited CD4bs‐targeting nanobodies, G36 and R27, when engineered into a triple tandem format with llama IgG2a‐hinge region and human IgG1‐constant region (G36×3‐IgG2a and R27×3‐IgG2a), neutralized 96% of a multiclade 208‐strain panel at geometric mean IC80s of 0.314 and 0.033 µg mL−1, respectively. Cryo‐EM structures of these nanobodies in complex with Env trimer revealed the two nanobodies to neutralize HIV‐1 by mimicking the recognition of the CD4 receptor. To enhance their neutralizing potency and breadth, nanobodies are linked to the light chain of the V2‐apex‐targeting broadly neutralizing antibody, CAP256V2LS. The resultant human‐llama bispecific antibody CAP256L‐R27×3LS exhibited ultrapotent neutralization and breadth exceeding other published HIV‐1 broadly neutralizing antibodies, with pharmacokinetics determined in FcRn‐Fc mice similar to the parent CAP256V2LS. Vaccine‐elicited llama nanobodies, when combined with V2‐apex broadly neutralizing antibodies, may therefore be able to fulfill anti‐HIV‐1 therapeutic and prophylactic clinical goals. [ABSTRACT FROM AUTHOR]

As the occurrences of cancer rise and the requirement for biotherapeutics increases, there is a growing demand for cost‐efficient methods to manufacture mammalian cell‐based biopharmaceuticals. Currently, the most cost‐efficient manufacturing solution is the cultivation of cells in perfusion mode. Perfusion allows for high cell densities of up to 200 million viable cells per mL (MVC mL−1) to be achieved, resulting in an increase in yield and volumetric productivity, which is the production of product per unit volume and time. However, culturing in perfusion mode requires large volumes of media, which is a significant expense in process development. Therefore, methods that allow rapid optimization of perfusion media are desirable to decrease operating costs and increase productivity. In this work, a quasi‐perfusion methodology using microwell plates (MWP) is used for media optimization to culture CHO‐S cells producing IgG1 monoclonal antibodies (mAb) known as trastuzumab, which have clinical applications treating HER2+ breast cancer. Results show blending glucose‐rich supplements and sodium butyrate with perfusion‐specific base media can lead to an 8‐fold increase in monoclonal antibody titre compared with traditional fed‐batch media. The original and optimized media were then scaled‐up to a custom made, mini bioreactor (MBR) running in perfusion mode. Two‐fold higher cell density is achieved in the MBR compared with the MWP, however, when normalizing for cell density, mAb productivity is comparable between the two methodologies. The combined MWP and 250 mL MBR methodology is an optimization tool that enables process development cost savings due to reduced volume of media utilization. [ABSTRACT FROM AUTHOR]