Addressing matrix effects in ligand-binding assays through the use of new reagents and technology.
BACKGROUND
Ligand-binding assays (LBAs) used in the quantification of biotherapeutics for pharmacokinetic determinations rely on interactions between reagents (antibodies or target molecule) and the biotherapeutic. Most LBAs do not employ an analyte extraction procedure and are susceptible to matrix interference. Here, we present a case study on the development of a LBA for the quantification of a PEGylated domain antibody where matrix interference was observed. The assay used to support the single ascending dose study was a plate-based electrochemiluminescent assay with a lower limit of quantification of 80 ng/mL. To meet sensitivity requirements of future studies, new reagents and the Gyrolab™ Workstation were evaluated.


RESULTS
Assay sensitivity improved nearly threefold in the final method utilizing new antibody reagents, a buffer containing blockers to human anti-animal antibodies, and the Gyrolab Workstation.


CONCLUSIONS
Experimental data indicate that all factors changed played a role in overcoming matrix effects.
Bristol-Myers Squibb, Route 206 & Province Line Rd, Princeton, NJ 08543, USA.
Shannon D
SD
Chilewski
Johanna R
JR
Mora
Carol
C
Gleason
Binodh
B
DeSilva
D000818:Animals; D001688:Biological Products / Q000032:analysis* / Q000493:pharmacokinetics / Q000494:pharmacology; D055808:Drug Discovery; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods*; D062069:Inventions; D008024:Ligands
D001688:Biological Products; D008024:Ligands