Effect of conditions of monoclonal antibody adsorption on antigen-binding activity

Tarakanova, Yu. N.; Dmitriev, D. A.; Massino, Yu. S.; Smirnova, M. B.; Segal, O.L.; Fartushnaya, O. V.; Yakovleva, D. A.; Kolyaskina, G. I.; Лавров, В. Ф.; Дмитриев, А. Д.

doi:10.1134/s0003683812050122

Cited by 6 publications

(4 citation statements)

References 18 publications

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“…Several factors that could be explored to lower detection limits by improving antigen–Ab binding are ionic strength, temperature, and pH. Neutral pH is commonly used for immunoaffinity assays, although research suggests that immobilized mAb to antigen binding capacity may improve as pH decreases to ∼3 . However, not all immune complexes are stable at low pH; further work would be required to investigate this possibility more fully, which is beyond the scope of this article.…”

Section: Resultsmentioning

confidence: 99%

Microchip immunoaffinity electrophoresis of antibody–thymidine kinase 1 complex

2015

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Thymidine kinase-1 (TK1) is an important cancer biomarker whose serum levels are elevated in early cancer development. We developed a microchip electrophoresis immunoaffinity assay to measure recombinant purified TK1 (pTK1) using an antibody that binds to human TK1. We fabricated poly(methyl methacrylate) microfluidic devices to test the feasibility of detecting antibody (Ab)-pTK1 immune complexes as a step towards TK1 analysis in clinical serum samples. We were able to separate immune complexes from unbound antibodies using 0.5X phosphate buffer saline (pH 7.4) containing 0.01% Tween-20, with 1% w/v methylcellulose that acts as a dynamic surface coating and sieving matrix. Separation of the antibody and Ab-pTK1 complex was observed within a 5 mm effective separation length. This method of detecting pTK1 is easy to perform, requires only a 10 μL sample volume, and takes just 1 minute for separation.

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Section: Resultsmentioning

confidence: 99%

Microchip immunoaffinity electrophoresis of antibody–thymidine kinase 1 complex

2015

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“…The electrolyte solution pH can also affect the antibody immobilization by physical adsorption . This dependence may be related to the differences in the charge and the degree of hydrophobicity of the variable regions of antibodies that may determine the orientation and conformational state of antibody molecules, which are dependent on the conditions of their adsorption . Figure shows the electrochemical impedance spectra for different pH values.…”

Section: Resultsmentioning

confidence: 99%

“…22 This dependence may be related to the differences in the charge and the degree of hydrophobicity of the variable regions of antibodies that may determine the orientation and conformational state of antibody molecules, which are dependent on the conditions of their adsorption. 23 Figure 4 shows the electrochemical impedance spectra for different pH values. As can be clearly observed, the pH has a major effect on the dynamic adsorption and also the amount adsorbed.…”

Section: ■ Introductionmentioning

confidence: 99%

Troponin T Immunosensor Based on Liquid Crystal and Silsesquioxane-Supported Gold Nanoparticles

et al. 2014

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A nanostructured immunosensor based on the liquid crystal (E)-1-decyl-4-[(4-decyloxyphenyl)diazenyl]pyridinium bromide (Br-Py) and gold nanoparticles supported by the water-soluble hybrid material 3-n-propyl-4-picolinium silsesquioxane chloride (AuNP-Si4Pic(+)Cl(-)) was built for the detection of troponin T (cTnT), a cardiac marker for acute myocardial infarction (AMI). The functionalized nanostructured surface was used to bind anti-cTnT monoclonal antibodies through electrostatic interaction. The immunosensor (ab-cTnT/AuNP-Si4Pic(+)Cl(-)/Br-Py/GCE) surface was characterized by microscopy techniques. The electrochemical behavior of the immunosensor was studied by cyclic voltammetry and electrochemical impedance spectroscopy. A calibration curve was obtained by square-wave voltammetry. The immnunosensor provided a limit of detection of 0.076 ng mL(-1) and a linear range between 0.1 and 0.9 ng mL(-1) (appropriate for AMI diagnosis).

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“…The pH and NaCl content can affect the charge on the epitope or paratope group and the available number of sites for the antigen-antibody reaction because they affect the hydrophobic interactions of the antigen and antibody (Reverberi & Reverberi, 2007;Tarakanova et al, 2012) and also influence the antibody titer and IC 50 of the ic-ELISA. Acetonitrile is used to extract AVM and IVM from animal-derived foods and can also influence the sensitivity of an ic-ELISA.…”

Section: Development and Optimization Of Ic-elisamentioning

confidence: 99%

Development of ic-ELISA and lateral-flow immunochromatographic assay strip for the simultaneous detection of avermectin and ivermectin

Xie

Kong

Liu

et al. 2017

Food and Agricultural Immunology

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A sensitive monoclonal antibody, 1H2, was generated in this study that simultaneously recognizes avermectin and ivermectin. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed, with 50% inhibitory concentrations of 0.491 ng/mL for avermectin and 0.770 ng/mL for ivermectin. A lateral-flow immunochromatographic assay (ICA) strip was also developed based on this monoclonal antibody for the semiquantitative and quantitative detection of avermectin and ivermectin. The semiquantitative (with the naked eye) analysis had visual limits of detection of 10 ng/mL for avermectin and 25 ng/mL for ivermectin, with cutoff values of 25 and 50 ng/mL, respectively. Using a strip scan reader, the quantitative results had calculated limits of detection of 1.3 ng/mL for avermectin and 2.9 ng/mL for ivermectin. In an analysis of raw milk samples, the recovery rates for the ic-ELISA ranged from 94% to 112% and for the ICA strip from 110% to 125%. Therefore, both methods are sensitive and effective for the on-site detection and rapid screening of samples, and are suitable for various other applications.

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Effect of conditions of monoclonal antibody adsorption on antigen-binding activity

Cited by 6 publications

References 18 publications

Microchip immunoaffinity electrophoresis of antibody–thymidine kinase 1 complex

Microchip immunoaffinity electrophoresis of antibody–thymidine kinase 1 complex

Troponin T Immunosensor Based on Liquid Crystal and Silsesquioxane-Supported Gold Nanoparticles

Development of ic-ELISA and lateral-flow immunochromatographic assay strip for the simultaneous detection of avermectin and ivermectin

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