High affinity human Fc specific monoclonal antibodies for capture kinetic analyses of antibody-antigen interactions.

Vishal Kamat, Candice Boutot, Ashique Rafique, Christian Granados, Jing Wang, Ashok Badithe, Marcela Torres, Ishita Chatterjee, Olav Olsen, William Olson, Tammy Huang

Analytical biochemistry 2022 Mar 01

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We recently demonstrated that capturing human monoclonal antibodies (hmAbs) using high affinity anti-human Fc (AHC) antibodies allows reliable characterization of antibody-antigen interactions. Here, we characterized six human Fc specific mouse monoclonal antibodies (mAbs) and compared their binding profiles with three previously characterized goat AHC polyclonal antibodies (pAbs), exhibiting properties of a good capture reagent. All six mouse AHC mAbs specifically bound with high affinity to the Fc region of hIgG1, hIgG2, hIgG4 and to 43 different hIgG variants, containing substitutions and/or mutations in the hinge and/or Fc region, that have been reported to exhibit modified antibody effector function and/or pharmacokinetics. Biacore sensor surfaces individually derivatized with mouse AHC mAbs exhibited >2.5-fold higher hIgG binding capacity compared to the three goat AHC pAb surfaces and reproducibly captured hIgG over 300 capture-regeneration cycles. The results of the capture kinetic analyses performed on 31 antibody-antigen interactions using surfaces derivatized with either of the two highest affinity AHC mAbs (REGN7942 or REGN7943) were in concordance with those performed using goat AHC pAb surfaces. Our data demonstrate that AHC mAbs such as REGN7942 and REGN7943 that have properties superior than the three goat AHC pAbs are highly valuable research reagents, especially to perform capture kinetic analyses of antibody-antigen interactions on optical biosensors. Copyright © 2021 Regeneron Pharmaceuticals. Published by Elsevier Inc. All rights reserved.

Citation

Vishal Kamat, Candice Boutot, Ashique Rafique, Christian Granados, Jing Wang, Ashok Badithe, Marcela Torres, Ishita Chatterjee, Olav Olsen, William Olson, Tammy Huang. High affinity human Fc specific monoclonal antibodies for capture kinetic analyses of antibody-antigen interactions. Analytical biochemistry. 2022 Mar 01;640:114455