Comparing ELISA and Surface Plasmon Resonance for Assessing Clinical Immunogenicity of Panitumumab

Abstract

Evaluation of the immunogenicity of panitumumab, a fully human anti-epidermal growth factor receptor mAb approved for use in colorectal cancer patients, led to the development of two separate immunoassays for the detection of anti-panitumumab Abs. The first immunoassay used a bridging ELISA capable of detecting 10 ng/ml positive control anti-panitumumab Ab. The ELISA incorporated an acid dissociation step to reduce drug interference and tolerated the presence of ∼100-fold molar excess of drug. During eight clinical trials, the ELISA detected developing Ab responses in 2 of 612 (0.3%) subjects. In one of the ELISA positive subjects, neutralizing Abs were detected using an epidermal growth factor receptor phosphorylation bioassay. The second immunoassay used a Biacore biosensor immunoassay format capable of detecting 1 μg/ml positive control Ab while tolerating the presence of equal molar amounts of drug. Although less sensitive and less tolerant to competing drug in the assay, the Biacore assay detected developing Ab responses in 25 of the 604 (4.1%) subjects. Additionally, the Biacore assay identified eight subjects who developed neutralizing Abs. Mouse mAbs with affinities ranging from 1.1 × 10−6 to 8.4 × 10−10 M were used to characterize both assay types. The ELISA was more sensitive for the detection of higher affinity mAbs and detected high-affinity mAbs in the presence of higher molar ratio of drug to mAb. The Biacore assay was more sensitive for detection of lower affinity mAbs and detected low affinity Abs in the presence of higher molar ratios of drug to mAb.

Induction of Human Anti-Human Antibody Responses (Ab2) after Application of a Humanized Lewis Y Carbohydrate Specific Antibody (Ab1): Connection of Prolonged Disease Stabilization with Ab3 Induction?

“…This finding warranted a side-by-side comparison of both techniques to assess the possibility of identifying false positive/false negative HAHA responders. Such approach was taken by Lofgren et al [27] and indeed they identified more HAHA positives samples when applying SPR technology.…”

Comparing Exponentially Weighted Moving Average and Run Rules in Process Control of Semiquantitative Immunogenicity Immunoassays

“…Assay control data were generated by three validated semiquantitative immunogenicity immunoassays each using a different previously described detection technology-electrochemiluminescence (ECL) (8), ELISA (9), and surface plasmon resonance (SPR) (10). Each immunogenicity immunoassay was used in the routine analysis of clinical samples.…”

Detection of cynomolgus monkey anti-protein XYZ antibody using immunocapture-LC/MS

“…Enzyme-linked immunosorbent assays (ELISA) and electrochemiluminescence (ECL) immunoassays [7] are among the most widely used platform for ADA detection. Other platforms that have been used include surface plasmon resonance and bio-layer interferometry which may be more suited for detection of low affinity ADA [8]. Typically, detection of ADA is followed by assessments of the magnitude (titer) of the ADA response and the in vitro neutralizing ability of ADA, especially in late-stage clinical studies.…”