Products/Services Used | Details | Operation |
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Proteins, Expression, Isolation and Analysis | … Louis, MO). Surrogate peptide VVSVLTVTHQDWLNGK was synthesized at Genscript (Piscataway, NJ). Streptavidin magnetic beads (1μm diameter.), TPCK trypsin and EZ-Link Sulfo-NHS-LC Biotinylation Kits were obtained from Thermo Scientific (Rockford, IL) | Get A Quote |
Although enzyme-linked immunosorbent assays and electrochemiluminescence (ECL) immunoassays are the most widely used platform for ADA detection, they may be compromised by drug interference. We describe here an alternate, free of drug interference, immunocapture-LC/MS methodology for detecting anti-protein XYZ antibody in cynomolgus monkey plasma. In our approach, ADA-protein XYZ complexes are captured by a mouse monoclonal anti-drug antibody on streptavidin magnetic beads and separated from monkey plasma by a magnet. After elution, ADA are digested with trypsin and detected by LC/MS using a surrogate peptide common to monkey IgG subclasses 1-4. The immunocapture-LC/MS assay was applied to support a toxicology ... More