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Novel methods to determine complement activation in human serum induced by the complex of Dezamizumab and serum amyloid P by Jianhong Ma; Qi Liu; John R. White is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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Lack of simple and robust methods to determine complement activation in human serum induced by antigen–antibody complexes is a major hurdle for monitoring therapeutic antibody drug quality and stability. Dezamizumab is a humanized IgG1 monoclonal antibody that binds to serum amyloid P component (SAP) for potential treatment of systemic amyloidosis. The mechanism of action of Dezamizumab includes the binding of SAP, complement activation through classical pathway, and phagocytosis; however, the steps in this process cannot be easily monitored. We developed two novel methods to determine Dezamizumab-SAP complex-induced complement activation. Complement component 3 (C3) depletion was detected by homogeneous time-resolved fluorescence (HTRF), and C3a desArg fragment, formed after the cleavage of C3 to yield C3a followed by removal of its C-terminal arginine residue, was determined using Meso Scale Discovery (MSD) technology. We found that the presence of both Dezamizumab and SAP was required for complement activation __via__ both methods. The optimal molar ratio of Dezamizumab:SAP was 6:1 in order to obtain maximal complement activation. The relative potency from both methods showed a

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Author

Jianhong Ma; Qi Liu; John R. White

Publisher

Elsevier BV

Published

2021

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)