IgG subclass specificity to C1q determined by surface plasmon resonance using Protein L capture technique

Rekha Patel; Alyssa Neill; Hongcheng Liu; Bruce Andrien

doi:10.1016/j.ab.2015.03.012

IgG subclass specificity to C1q determined by surface plasmon resonance using Protein L capture technique

Anal Biochem. 2015 Jun 15:479:15-7. doi: 10.1016/j.ab.2015.03.012. Epub 2015 Mar 19.

Authors

Rekha Patel¹, Alyssa Neill², Hongcheng Liu², Bruce Andrien²

Affiliations

¹ Product Characterization, Alexion Pharmaceuticals, Inc, USA. Electronic address: [email protected].
² Product Characterization, Alexion Pharmaceuticals, Inc, USA.

PMID: 25797348
DOI: 10.1016/j.ab.2015.03.012

Abstract

Recombinant monoclonal antibodies (mAbs) have become an important category of biological therapeutics. mAbs share the same structures and biological functions as endogenous IgG molecules. One function is complement-dependent cytotoxicity (CDC) initiation by binding of C1q. Traditionally, ELISA methods have been utilized to measure C1q binding. A new robust capture method was established in this study to measure the binding affinity of C1q to antibodies by surface plasmon resonance (SPR). The utility of this method was demonstrated by determination of the difference in IgG subclass specificity of C1q binding.

Keywords: C1q; Complement dependent cytotoxicity; Monoclonal antibodies; Surface plasmon resonance.

MeSH terms

Animals
Antibodies, Monoclonal / analysis
Antibodies, Monoclonal / immunology
Bacterial Proteins / chemistry
CHO Cells
Complement C1q / immunology*
Cricetulus
Humans
Immobilized Proteins / chemistry
Immunoglobulin G / analysis
Immunoglobulin G / immunology*
Peptococcus / chemistry
Recombinant Proteins / analysis
Recombinant Proteins / immunology
Surface Plasmon Resonance / methods*

Substances

Antibodies, Monoclonal
Bacterial Proteins
Ig L-binding protein, Peptostreptococcus
Immobilized Proteins
Immunoglobulin G
Recombinant Proteins
Complement C1q