Impact of antibody subclass and disulfide isoform differences on the biological activity of CD200R and βklotho agonist antibodies
| Autor: | Brian Mingtung Chan, Robert Y.-T. Chou, Ognjen Grujic, Stephanie A Grauer, Kevin Graham, Christy Ann Thomson, Yang Li, Jennifer Weiszmann, Laura Sekirov, Jennitte Stevens, Ian Foltz, Kathy Manchulenko, Anita Badh, Thomas M. Dillon |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Agonist medicine.drug_class Biophysics Receptors Cell Surface CHO Cells Monoclonal antibody Biochemistry 03 medical and health sciences Structure-Activity Relationship 0302 clinical medicine Cricetulus Cell surface receptor Orexin Receptors parasitic diseases medicine Animals Protein Isoforms Avidity Disulfides Receptor Molecular Biology Klotho Proteins biology Antibodies Monoclonal Membrane Proteins Biological activity Cell Biology Primary and secondary antibodies Molecular biology 030104 developmental biology 030220 oncology & carcinogenesis Antigens Surface biology.protein Antibody Epitope Mapping |
| Zdroj: | Biochemical and biophysical research communications. 486(4) |
| ISSN: | 1090-2104 |
| Popis: | Agonism of cell surface receptors by monoclonal antibodies is dependent not only on its ability to bind the target, but also to deliver a biological signal through receptors to the cell. Immunoglobulin G2 antibodies (IgG2s) are made up of a mixture of distinct isoforms (IgG2-A, -B and A/B), which differ by the disulfide connectivity at the hinge region. When evaluating panels of agonistic antibodies against CD200 receptor (CD200R) or βklotho receptor (βklotho), we noticed striking activity differences of IgG1 or IgG2 antibodies with the same variable domains. For the CD200R antibody, the IgG2 antibody demonstrated higher activity than the IgG1 or IgG4 antibody. More significantly, for βklotho, agonist antibodies with higher biological activity as either IgG2 or IgG1 were identified. In both cases, ion exchange chromatography was able to isolate the bioactivity to the IgG2-B isoform from the IgG2 parental mixture. The subclass-related increase in agonist activity was not correlated with antibody aggregation or binding affinity, but was driven by enhanced avidity for the CD200R antibody. These results add to the growing body of evidence that show that conformational differences in the antibody hinge region can have a dramatic impact on the antibody activity and must be considered when screening and engineering therapeutic antibody candidates. The results also demonstrate that the IgG1 (IgG2-A like) or the IgG2-B form may provide the most active form of agonist antibodies for different antibodies and targets. |
| Databáze: | OpenAIRE |
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