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Considering the limitations of well-known traditional detection techniques, innovative research studies have focused on the development of new sensors to offer label-free, highly sensitive, real-time, low-cost, and rapid detection for biomolecular interactions. In this study, we demonstrate immunoglobulin G (IgG) detection in aqueous solutions by using real-time and label-free kinetic measurements of the Interferometric Reflectance Imaging Sensor (IRIS) system. By performing kinetic characterization experiments, the sensor's performance is comprehensively evaluated and a high correlation coefficient value (>0.94) is obtained in the IgG concentration range of 1-50 μg/mL with a low detection limit (0.25 μg/mL or 1.67 nM). Moreover, the highly sensitive imaging system ensures accurate quantification and reliable validation of recorded binding events, offering new perspectives in terms of direct biomarker detection for clinical applications., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The immobilized metal-affinity chromatography (IMAC) has gained significant interest as a widespread separation and purification tool for therapeutic proteins, nucleic acids and other biological molecules. The enormous potential of IMAC for proteins with natural surface exposed-histidine residues and for recombinant proteins with histidine clusters. Cryogels as monolithic materials have recently been proposed as promising chromatographic adsorbents for the separation of biomolecules in downstream processing. In the present study, IMAC cryogels have been synthesized and utilized for the adsorption and separation of immunoglobulin G (IgG) from IgG solution and whole human plasma. For this purpose, Cu(II)-ions were coupled to poly(hydroxyethyl methacrylate) PHEMA using poly(ethylene imine) (PEI) as the chelating ligand. In this study the cryogels formation optimized by the varied proportion of PEI from 1% to 15% along with different amounts of Cu (II) as chelating metal. The prepared cryogels were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The [PHEMA/PEI]-Cu(II) cryogels were assayed for their capability to bind the human IgG from aqueous solutions. The IMAC cryogels were found to have high affinity toward human IgG. The adsorption of human IgG was investigated onto the PHEMA/PEI cryogels with (10% PEI) and the concentration of Cu (II) varied as 10, 50, 100 and 150 mg/L. The separation of human IgG was achieved in one purification step at pH7.4. The maximum adsorption capacity was observed at the [PHEMA/PEI]-Cu(II) (10% PEI) with 72.28 mg/g of human IgG. The purification efficiency and human IgG purity were investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this study, thiophilic cryogels were prepared by two different approaches and they were used in purification of IgG from aqueous solutions and human plasma. In the first approach, poly(2-hydoxyethyl methacrylate) [PHEMA] cryogel disks were prepared. The PHEMA cryogel disks were activated by divinylsulfone (DVS) and 2-mercapto ethanol was attached. In the second approach, poly(2-hydroxyethyl methacrylate-etylene glycol dimethacrylate) [P(HEMA-EGDMA)] beads were synthesized and 2-mercapto ethanol was attached to the beads as a thiophilic ligand. In order to increase the surface area, P(HEMA-EGDMA)/PHEMA composite cryogel disks were prepared by embedding the P(HEMA-EGDMA) beads into PHEMA cryogels. Both the thiophilic PHEMA (T-PHEMA) cryogel disks and the thiophilic P(HEMA-EGDMA)/PHEMA [T-P(HEMA-EGDMA)/PHEMA] composite cryogel disks were characterized by Fourier transform infrared spectroscopy, surface area measurements, elemental analysis, swelling tests and scanning electron microscopy. The effects of salt concentration, pH, temperature, initial IgG concentration were analysed for IgG adsorption from aqueous solutions in batch mode. When lyotropic salt, Na2SO4, was used, the adsorption capacity was 27.5 mg/g and 68.7 mg/g for T-PHEMA and the T-P(HEMA-EGDMA)/PHEMA composite cryogel disks, respectively. 1.0 M NaCl was used as desorption agent. The change in adsorption capacity was not remarkable for a repetition of adsorption-desorption cycle ten times. Both the thiophilic cryogel disks and the thiophilic composite cryogel disks were also successfully used for IgG isolation from human plasma. The purity assayed by SDS-PAGE was 89%. The adsorption capacities were 74.8 mg/g and 137.4 mg/g in human plasma samples for the T-PHEMA cryogel disks and the T-P(HEMA-EGDMA)/PHEMA composite cryogel disks, respectively., (Copyright © 2013 Elsevier B.V. All rights reserved.)