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Dear Editor,A 62-year-old woman was admitted to our hospital because of edema of both lower limbs for more than 3 months and oliguria for 1 week. The patient’s laboratory test results were listed i...

The aim of this study was to investigate neonatal Fc receptor (FcRn) concentration developmental pharmacology in adult and pediatric subjects using minimal physiologically-based pharmacokinetic (mPBPK) modelling. Three types of pharmacokinetic (PK) data for three agents (endogenous/exogenous native IgG, bevacizumab and palivizumab) were used. The adult group contained six subjects with weights from 50 to 100 kg. For pediatric subjects, seven age groups were assumed, with five subjects each having the weight of 95%, 75%, 50%, 25% and 5% percentile of the population. A first evidence-based rating system to evaluate the quality of the source data used to derive pediatric-specific mPBPK model parameter was proposed. A stepwise approach was used to examine the best combination of age/weight effect on the parameters of the mPBPK model in adult and pediatric subjects. IgG synthesis rate (K(syn)), extravasation rate (ER) and FcRn were fitted simultaneously to the PK of bevacizumab and native-IgG in both adult and pediatric. All fitting showed good fits based on the graphs and the coefficient of variation of the fitted parameters (

We report the case of a French woman with acquired von Willebrand syndrome who presents recurrent subarachnoid and intra-cerebral hemorrhage since 2012. She had no family or personal bleeding history. In the biologic explorations, APTT was abnormally high with no anticoagulant drugs (it was normal, historically). Two monoclonal IgG and IgM kappa proteins were detected without any lymphoproliferative disorder. Intravenous infusion of immunoglobulin is very effective in AVWS with immunoglobulin G monoclonal gammapathie of undetermined significance. We had a satisfactory correction of coagulation factors for about 30 days. The exploration of APTT is surely essential for the diagnosis and treatment.

Proliferative glomerulonephritis with monoclonal immunoglobulin G deposits (PGNMID) is a newly identified and rare form of glomerulonephritis which is characterized by endocapillary proliferative or membranoproliferative with monoclonal deposit stain for IgG and a single light chain. We describe the case of a 63-year-old woman with type 2 diabetes who was considered to have PGNMID.

Glomerulopathy due to dysproteinemia can have a wide spectrum of pathologic and clinical features based on specific characteristics of the abnormal protein and the response induced within the parenchymal tissue. Monoclonal immunoglobulin G (IgG) deposition can manifest as a different glomerular disease. Proliferative glomerulonephritis (GN) with monoclonal IgG deposits (PGNMID) is a unique entity mimicking immune complex GN that does not conform to any of those subtypes. IgG monoclonal granular deposition in the glomeruli with a pattern similar to immune complex disease suggested by C3 and C1q deposition should prompt consideration of PGNMID. Literature is scarce in terms of recurrence of disease in renal allografts. In this article we present the clinical-pathologic features of three cases of PGNMID in the renal allograft showing the variable course and manifestation of the disease.

Proliferative glomerulonephritis occurring as a consequence of monoclonal glomerular deposits of IgG is uncommon. It is a form of renal involvement in monoclonal gammopathy that mimics immune complex glomerulonephritis. Here, we report the first series of proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) from the Indian subcontinent highlighting use of light chain immunofluorescence (IF) in routine renal biopsy interpretation. We retrieved 6 patients diagnosed as proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) out of 160 biopsies (3.7%) with membranoproliferative patterns over 5 1/2 years (2009–2014), one of whom had recurrence 6 months post-renal transplant. Four (67%) patients presented with rapidly progressive renal failure and two (33%) with nephrotic syndrome. None of these patients had overt multiple myeloma. The predominant histologic pattern was membranoproliferative with all the biopsies showing IgG3 Kappa deposits on IF. The deposits were primarily subendothelial on electron microscopy.

International audience; Le STUDIUM conference was held November 24-25, 2016 in Tours, France as a satellite workshop of the 5(th) meeting of the French GDR 3545 on "G Protein-Coupled Receptors (GPCRs) - From Physiology to Drugs", which was held in Tours during November 22-24, 2016. The conference gathered speakers from academia and industry considered to be world leaders in the molecular pharmacology and signaling of GPCRs, with a particular interest in the development of therapeutic GPCR antibodies (Abs). The main topics were new advances and challenges in the development of antibodies targeting GPCRs and their potential applications to the study of the structure and function of GPCRs, as well as their implication in physiology and pathophysiology. The conference included two sessions, with the first dedicated to the recent advances in methodological strategies used for GPCR immunization using thermo-stabilized and purified GPCRs, and the development of various formats of Abs such as monoclonal IgG, single-chain variable fragments and nanobodies (Nbs) by in vitro and in silico approaches. The second session focused on GPCR Nbs as a "hot" field of research on GPCRs. This session started with discussion of the pioneering Nbs developed against GPCRs and their application to structural studies, then transitioned to talks on original ex vivo and in vivo studies on GPCR-selective Nbs showing promising therapeutic applications of Nbs in important physiological systems, such as the central nervous and the immune systems, as well as in cancer. The conference ended with the consensus that Abs and especially Nbs are opening a new era of research on GPCR structure, pharmacology and pathophysiology.

Proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) is a recently described entity. It is featured by glomerular nonorganized monoclonal immunoglobulin G deposits. Monoclonal IgG deposits are associated with glomerular proliferative lesions, mimicking different types of immune-complex glomerulonephritis. We report two classic cases of PGNMID which fulfilled all criteria of this disease. We conclude that recognition of proliferative glomerulonephritis with monoclonal IgG deposits requires routine immunostaining for light chain and IgG isotype.

A male patient is presented with long-lasting paraproteinemia of monoclonal IgG λ, who suffered from recurrent, and until the last one, mostly reversible episodes of dermatoneuro syndrome, described exclusively in scleromyxedema. The skin biopsy revealed λ-light chain amyloid deposition instead of changes typical for scleromyxedema. Systemic AL amyloidosis was diagnosed post mortem since the patient had no clinical signs of any other organ impairment except skin and brain. Neuropathology is described and possible etiopathogenesis of brain involvement is considered.

Contains fulltext : 167798.pdf (Publisher’s version ) (Closed access) Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies.

In the diagnosis of atherosclerosis, detailed evaluation of biomarkers related to its lesion formation is desired for estimation of its progression rate. In our previous proteomic studies of atherosclerosis mice, the protein level of thrombospondin-4 (TSP4) in the aorta, but not in plasma, elevated relatively with atherosclerotic plaque formation. Therefore, we supposed that TSP4 would be a potential biomarker for diagnostic imaging of atherosclerotic progression. Immunoglobulin G (IgG) has been widely used as a basic molecule of imaging probes providing images specific to their target biomolecules, owing to the antigen-antibody reaction. Therefore, we first developed anti-TSP4 monoclonal IgG radiolabeled with 99mTc (99mTc-TSP4-mAb). 99mTc-TSP4-mAb showed higher accumulation in atherosclerotic aortas of apoE−/− mice (atherosclerotic model mice); however, we found that the non-targeted monoclonal IgG radiolabeled with 99mTc also showed similar distribution in atherosclerotic aortas of apoE−/− mice. IgG has also known to accumulate nonspecifically in the immunological disease such as inflammatory arthritis. However, the accumulation mechanism of IgG has still been unclear in detail. In this chapter, we would like to introduce recent topics on atherosclerotic imaging, focused on our work exploring the accumulation mechanisms of IgG in atherosclerotic lesions, and elucidating the usefulness of radiolabeled IgG images in the diagnosis of atherosclerosis.

Proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) is a rare and recently identified disease with a poor prognosis irrespective of the treatment. Recently, the possibility of recurrent or de novo PGNMID after kidney transplantation has been reported, which is associated with a better prognosis compared to PGNMID on native kidneys. Nevertheless, at present, due to the very few cases of recurrent PGNMID diagnosed, there is no proven effective treatment. Here, we report a case of recurrent PGNMID successfully treated with plasmapheresis, steroids and mycophenolate mofetil. Our report suggests that plasmapheresis might be a valid therapeutic option to treat recurrent PGNMID.

SummaryHuman cytomegalovirus (CMV) is a common but difficult to treat infection of immunocompromised patients. MSL-109 is a human monoclonal IgG isolated from a CMV seropositive individual that recognizes the viral glycoprotein H (gH) surface antigen complexes that mediate entry. Although MSL-109 blocks CMV infection in vitro, it lacked sufficient efficacy in human trials, and CMV isolated from treated patients suggested the evolution of MSL-109 resistance. To understand how CMV escapes MSL-109, we characterized a MSL-109-resistant CMV strain. Our results elucidate a nongenetic escape mechanism in which the antibody is selectively taken up by infected cells and incorporated into assembling virions in a dose-dependent manner. The resistant virus then utilizes the Fc domain of the incorporated antibody to infect naive nonimmune cells. This resistance mechanism may explain the clinical failure of MSL-109, illustrate a general mechanism of viral antibody escape, and inform antiviral vaccine and therapeutic development.

Membranous-like glomerulopathy with masked IgG-κ deposits (MGMID) is a novel entity requiring antigen retrieval on formalin-fixed paraffin-embedded tissue to detect the immunoglobulin by immunofluorescence. MGMID is clinically distinct from other glomerulopathies with non-organized monoclonal IgG deposits, although the source of the kappa-restricted IgG is uncertain. Careful examination including ultrastructural analysis is essential for identifying diseases such as MGMID that may be misclassified by routine methods and ultimately require alternative techniques for accurate diagnosis.

A robust cation-exchange chromatofocusing method was developed for the routine analysis of a recombinant humanized monoclonal IgG antibody. We compare the chromatofocusing method to the conventional cation-exchange chromatography (CEX) employing a salt gradient and show clear advantages of chromatofocusing over CEX. We demonstrate the suitability of the present chromatofocusing method for its intended purpose by testing the validation characteristics. To our knowledge, this is the first chromatofocusing method developed for the routine analysis of monoclonal antibody charge species.

This paper describes a method for the purification of monoclonal antibodies (rat anti-2,4-dinitrophenyl IgG: IgG(DNP); and mouse antidigoxin IgG: IgG(Dgn)) from ascites fluid. This procedure (for IgG(DNP)) has three steps: (i) precipitation of proteins heavier than immunoglobulins with ammonium sulfate; (ii) formation of cyclic complexes of IgG(DNP) by causing it to bind to synthetic multivalent haptens containing multiple DNP groups; (iii) selective precipitation of these dimers, trimers, and higher oligomers of the target antibody, followed by regeneration of the free antibody. This procedure separates the targeted antibody from a mixture of antibodies, as well as from other proteins and globulins in a biological fluid. This method is applicable to antibodies with a wide range of monovalent binding constants (0.1 microM to 0.1 nM). The multivalent ligands we used (derivatives of DNP and digoxin) isolated IgG(DNP) and IgG(Dgn) from ascites fluid in yields of >80% and with >95% purity. This technique has two advantages over conventional chromatographic methods for purifying antibodies: (i) it is selective for antibodies with two active Fab binding sites (both sites are required to form the cyclic complexes) over antibodies with one or zero active Fab binding sites; (ii) it does not require chromatographic separation. It has the disadvantage that the structure of the hapten must be compatible with the synthesis of bi- and/or trivalent analogues.

We report a 5-month-old Amish girl with atypical recombinase-activating gene (RAG)-deficient severe combined immunodeficiency disease. There was a lys992glu RAG-1 substitution leading to impaired RAG activity. Immunological studies revealed mildly decreased CD3+ T cells, markedly decreased CD4+ T cells, and oligoclonal T-cell receptor-Vβ T cells. B cells were markedly decreased but serum immunoglobulin levels were normal with monoclonal IgG and IgM antibodies.

First description of a case of early myeloma with monoclonal IgG, type kappa, anomaly plus Bence-Jones proteinuria and Fanconi's syndrome in a 67-year-old woman. Characteristic crystalline inclusion bodies were found in plasma cells of bone marrow and renal tissue.

A panel of four natural human monoclonal IgG antibodies derived from B lymphocytes isolated from regional draining lymph nodes of cancer patients has been developed and characterized. The four human antibodies are termed, RM1, RM2, RM3, and RM4. The immunoreactivity of this panel of four human antibodies is restricted to tumor cells. Individually, these human MAbs show tumor targeting and are effective in inhibiting tumor growth in nude mouse xenograft models. When used in combination the antibodies show an additive effect in slowing down the progression of tumors in xenograft models suggesting that cocktails of antibodies may be useful in the clinic.

An IgG mouse monoclonal antibody (10F05) against polyethylene glycol has been generated. The antibody reacts with PEG regardless of the linker used for PEG attachment, and is able to recognize a PEGylated peptide in plasma at concentrations as low as 3 pg/mL. The antibody is readily purified in substantial quantities. The PEG IgG will find significant utility in the sensitive detection of PEG derivitives during the pharmacokinetic characterization of PEGylated compounds.

This research was conducted to evaluate the changes in allergenicity of shrimp by physical treatments using competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). Shrimp was subjected to physical treatments such as high hydrostatic pressure (HHP), sonication, autoclave and microwave. Heat-stable protein (HSP) purified from raw shrimp was used as a major allergen. The binding ability of monoclonal IgG and shrimp-allergic patients` IgE to HSP treated with HHP decreased, increasing the pressure up to 400 MPa. Especially, it became less than 50% at 400 MPa. The binding ability of mAb to HSP treated with sonication (10, 20, 30 and 60 min) decreased with the treated time. Especially, it became less than 60% with the treatment for 60 min. The allergenicity change of HSP treated with autoclave and microwave little decreased. The binding ability of mAb to HSP during the treatment for 20 min became more than 70%. The results suggest that allergenicity of HSP in raw shrimp be more easily lost by HHP or sonication treatment than by autoclave or microwave treatment.

Summary The features of GPs with Congo red-negative, non-cryoglobulinaemic deposits have been surveyed. Thesedisorders are rare and clinically heterogeneous andtheir pathogenesis is unclear. There is controversyconcerning the nomenclature of Congo-red negativeglomerular fibrilloses. A practical and reproducibleapproach is to classify these GPs on the basis of thesubstructure and arrangement of the deposits assessedat conventional electron microscopic magnifications.Accordingly, FGN and ITG can be identified. FGN ischaracterized by randomly arranged microfibrils (dia-meter: 12–30nm) composed of subclass-restrictedpolyclonal IgG and a low incidence of underlyingsystemic disease. In contrast, ITG is characterized byparallel bundles of microtubules (diameter: 10–90nm)composed mainly of monoclonal IgG and a highincidence of associated lymphoproliferative disease ormonoclonal gammopathy. The finding of Congo red-negative organized deposits on renal biopsy shouldprompt a careful search and follow-up for mono-clonal gammopathy, cryoglobulins and haemopoieticmalignancy.

Vibratory angio-oedema is a rare form of physical urticaria characterized by pruriginous weals and angio-oedema at the site of exposure to vibration. Severe treatment-resistant disease can occur, and is associated with significant disability. Therapy with omalizumab, a monoclonal IgG anti-IgE antibody, has been shown to be successful in several types of physical urticaria. We report a patient with vibratory angio-oedema for whom all standard treatments for urticaria, including omalizumab, failed to show a clinical benefit. Finally, ketotifen was tried, and unexpectedly reduced symptoms significantly. Ketotifen may thus represent a therapeutic option in patients with treatment-resistant vibratory angio-oedema.

Clostridium difficile infection has emerged as a growing worldwide health problem. The colitis of Clostridium difficile infection results from the synergistic action of C. difficile secreted toxins A and B upon the colon mucosa. A human monoclonal IgG anti-toxin has demonstrated the ability in combination therapy to reduce mortality in C. difficile challenged hamsters. This antibody is currently in a clinical trial for the treatment of human Clostridium difficile infection. More than one group of investigators has considered using polyclonal bovine colostral antibodies to toxins A and B as an oral passive immunization. A significant proportion of the healthy human population possesses polyclonal antibodies to the Clostridium difficile toxins. We have demonstrated that polyclonal IgA derived from the pooled plasma of healthy donors possesses specificity to toxins A and B and can neutralize these toxins in a cell-based assay. This suggests that secretory IgA prepared from such pooled plasma IgA may be able to be used as an oral treatment for Clostridium difficile infection.

A 69-year-old man presented with severe pain in hands and feet related to necrosis. Extensive explorations revealed type I cyroglobulinemia associated to monoclonal IgG kappa. Unfortunately he died a few months after.

A 78-year-old woman developed acute-onset nephrotic syndrome. A renal biopsy showed mild mesangial proliferative glomerulonephritis. Immunofluorescence studies revealed granular IgG3- λ deposits within the mesangial area and along the glomerular capillary walls. Electron microscopy showed mesangial and subendothelial granular electron-dense deposits. The pattern of deposition was predominantly mesangial. Serum or urine monoclonal proteins were not detected. Middle-dose steroid therapy induced a rapid remission of nephrotic syndrome. We consider that this is the first case of steroid-responsive nephrotic syndrome due to an extremely rare glomerular disease, proliferative glomerulonephritis with monoclonal IgG deposits associated with pure mesangial proliferative features.

A residual IgG line is frequently observed inside the monoclonal IgG line on immunoelectrophoresis of IgG myeloma serum in reaction with anti-whole human serum. We found that the residual IgG line was observed exclusively in serum from IgG-λ type M-proteinemia when a goat antiserum to whole human serum was employed. However, with a horse antiserum to whole human serum, the residual IgG line was observed in sera from both the IgG-λ and κ type M-proteinemias. These phenomena led us to an idea that the appearance of the residual IgG line resulted from reaction with anti-κ or λ antibodies present in the anti-whole human serum antisera. Investigations of sera from a number of IgG M-proteinemia by anti-κ and λ antisera, and absorption tests with partially purified BJP of both types indicated that the occurrence of the residual IgG line is due to the reaction of polyclonal IgG with anti-κ or λ chain antibody present in the anti-whole human serum.

Purpose To detect and characterize the aggregation of therapeutic monoclonal antibodies in undiluted biological fluids. Methods Fluorescently labeled subvisible IgG aggregates formed by applying either heat stress or by pH-shift were investigated immediately after addition to human serum, and after 24 h. Unstressed and stressed IgG formulations were analyzed by fluorescence single particle tracking, confocal laser scanning microscopy and flow cytometry. Results Unstressed formulations remained free from subvisible aggregates in serum, whereas heat-stressed and pH-shift stressed formulations showed dissimilar aggregation behaviors. The aggregation profile of the heat-stressed formulation diluted in serum remained practically the same as the one diluted in buffer, even after the 24 h incubation period. The pH-shift stressed formulation had strikingly smaller and more numerous subvisible aggregates immediately after dilution in serum compared to buffer. These aggregates became noticeably larger in both diluents after 24 h, but in serum they appeared to be formed by other types of constituents than the labeled protein itself. Conclusion These results show that subvisible therapeutic protein aggregates may undergo changes in number, type and size distribution upon contact with human serum. This emphasizes the importance of analytical strategies for monitoring aggregation in undiluted biological fluids. Electronic supplementary material The online version of this article (doi:10.1007/s11095-012-0749-x) contains supplementary material, which is available to authorized users.

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The prevalence of peripheral blood B cells secreting antibodies reacting with the F(ab')2 fragment of monoclonal IgG was studied in five patients with multiple myeloma (MM), nine patients with monoclonal gammopathy of undetermined significance (MGUS) and six healthy controls. An enzyme-linked immunospot assay allowed direct visualization of antibody producing B cells. All patients had B cells producing antibodies to autologous or allogeneic monoclonal IgG. Autoreactive cells were found more frequently than alloreactive cells in seven out of nine patients with MGUS and three out of four patients with MM. The same frequency of alloreactive cells in the patient groups was detected in healthy individuals. These findings show the existence of B cells producing anti-idiotypic antibodies which could be a part of an idiotypic network in monoclonal gammopathies.

To shed further light on plasmin-IgG interactions a simple procedure is described that permitted 48 monoclonal IgG isolates from human serum to be profiled for their susceptibility to plasmic cleavage. In addition to anodal Fc and cathodal Fab fragments, combined immunoelectrophoresis electrophoresis revealed transient anodal banding, as well as Fab-fragment subcleavage in many of the IgG subclass-1 isolates. The subcleavage of released Fab fragments, which bear the idiotype determinants, points to a possible ancillary role of plasmin in "idiotype processing" leading to immunoregulatory antiidiotype networks. The cleavage of IgG by endogenous plasmin also points to a possible active role of plasmin in the "steady-state" metabolism of IgG.

The POEMS syndrome is an infrequently reported multisystem disorder which presents usually as an obscure polyneuropathy, with almost all cases reported in Japan. A 64 yr old caucasian man presented with a 12 mth history of a severe sensorimotor neuropathy in association with dermato-endocrine features. Detection of a monoclonal IgG kappa paraprotein and mixed osteosclerotic/lytic bone lesions consistent with a plasma cell dyscrasia led to diagnosis of the POEMS syndrome. Unique ultrastructural features were present on sural nerve biopsy in addition to the unusual association with monoclonal kappa-light chain. This case illustrates that the POEMS syndrome may also occur in caucasian subjects.

Monoclonal IgG rheumatoid factors (RFs) are identified using a novel immunoblot assay which detects RFs that bind to SDS-denatured Fc on nitrocellulose. Recognition of these self-associating antibodies is by the use of F(ab′) 2 fragments of light chain-specific antisera. In this way, IgG RFs can be easily identified and the precise binding characteristics to different isotypes of IgG, or other antigens, further specified. The assay can also be used to detect other classes of RFs such as IgM RFs. Although less sensitive than the standard ELISA, the use of this immunoblot RF assay (IRFA) will identify IgG RFs and their target antigen with precision.

Summary Cholinergic urticaria is one of the more common physical urticarias. Although it is often fairly mild, severe treatment-resistant disease may occur, with significant associated disability. Omalizumab, a monoclonal IgG anti-IgE antibody licensed for use in severe asthma, has recently been used successfully in several types of urticaria, including in one case of cholinergic urticaria. This paper reports a patient with severe cholinergic urticaria, unresponsive to antihistamines and multiple other treatments, whose disease was also unresponsive to omalizumab.

We report a 74-year-old woman with a slowly progressive sensory motor axonal neuropathy and a monoclonal IgG-kappa that bound to a 68-kd axonal protein identified as the low molecular weight neurofilament protein. The sera of control subjects and disease controls did not bind to neurofilament protein.

The roles of monoclonal antibody affinity and treatment time of (+)-methamphetamine-induced pharmacological effects in rats were studied using two anti-(+)-methamphetamine monoclonal antibodies. These studies tested the preclinical protective effects of monoclonal antibody antagonists in (+)-methamphetamine overdose and pretreatment scenarios. The higher affinity antibody (mAb6H4; KD=11 nM for (+)-methamphetamine) more effectively antagonized (+)-methamphetamine-induced behavioral effects (distance and rearing) than the low affinity antibody (designated mAb6H8; KD=250 nM) and had a longer duration of action. Both antibodies more effectively reduced (+)-methamphetamine effects in the overdose model than in the pretreatment model. (+)-Methamphetamine pharmacokinetic studies showed the mAb6H4 significantly reduced brain concentrations over time in both models. However, while mAb6H4 immediately reduced brain concentrations in the overdose model, it did not prevent the initial distribution of (+)-methamphetamine into the brain in the pretreatment model. Thus, anti-(+)-methamphetamine monoclonal antibody affinity and administration time (relative to (+)-methamphetamine dosing) are critical determinants of therapeutic success.

A surface plasmon resonance (SPR) method for monitoring the concentration of the chaperone DnaK and its relation to physiological stress response in a recombinant Escherichia coli strain subjected to heat shock is described. The DnaK protein, an abundantly occurring representative of the heat-shock proteins, was used as a marker of physiological stress. The SPR biosensor instrument was used for label-free immunoaffinity detection directly in cell culture lysates using an anti-DnaK monoclonal IgG antibody immobilized on the sensor surface. The SPR method provides a fast response (

Streptococcus mutans is known to be a prime etiologic agent for the initiation and progression of human dental caries. Rapid, accurate, and quantitative detection of S. mutans will help us better understand the pathogenic mechanisms of dental caries and will help to develop methods for caries diagnosis and risk assessment. This study describes the development of three highly species-specific monoclonal IgG antibodies against S. mutans. The antibodies were used to develop a number of methods that quantitatively detect S. mutans in less

A sensitive test for the presence of D-positive fetal red blood cells (RBCs) in the maternal circulation of D-negative women has been developed. It was used to investigate the possibility that the occasional failure in preventing alloimmunization might be due to the administration of inadequate amounts of prophylactic anti-D Rh immune globulin. The standard dose in Australia contains 125μg of antibody, and can suppress immunization by an estimated 6 mL of packed D-positive RBCs. A fetomaternal hemorrhage (FMH) of this volume is detectable in the maternal circulation as approximately 0.25 percent of the total RBCs. Our test utilizes a commercially available human monoclonal IgG anti-D that has been biotinylated and used with a dye-conjugated streptavidin. Flow cytometry is used to quantitate fluorescing D-positive RBCs. To date, 2,288 tests have been performed on blood samples from D-negative women attending local antenatal clinics or at the time of delivery. Evidence for an FMH has been obtained in six cases (0.26%). In one case, the FMH was only 0.1 percent, and in another (confirmed by the Kleihauer-Betke method), fetal cells constituted only 0.2 percent. Additional Rh immune globulin was not given to these patients. In the other four cases, the D-positive fetal cells were estimated to be 0.7, 0.5, 0.5, and 0.4 percent, and additional prophylactic Rh immune globulin was administered. Although the prevalence of FMH is low, screening D-negative women at risk of alloimmunization has proved to be simple, fast, and inexpensive.

We report the case of a woman, who initially presented with an IgG kappa-type monoclonal gammopathy. An IgD-secreting myeloma was diagnosed 2 years later. The patient died of severe renal failure and infection. The discrepancy between the immunoglobulin concentration estimated from the electrophoresis pattern and the immunonephelometric measurement of IgG, IgA and IgM led us to investigate the existence of an IgD myeloma. It was of the IgD kappa type and had electrophoretic characteristics identical to the initial monoclonal IgG. A common clonal origin for these two immunoglobulins is discussed.