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- Title
Development, Validation and Application of a Bridging ELISA for Detection of Antibodies against GQ1001 in Cynomolgus Monkey Serum.
- Authors
Liu, Tingting; Sun, Yajun; Deng, Xiaojie; Shi, Lili; Chen, Wenyi; Fang, Wenjing; Wu, Junliang; Fan, Xiaotian; Chen, Xiaoqiang; Sun, Jianhua; Qin, Gang; Gong, Likun; Qin, Qiuping
- Abstract
Immunogenicity is a major issue associated with the PK, efficacy, and safety evaluation of therapeutic protein products during pre-clinical and clinical studies. A multi-tiered approach consisting of screening, confirmatory, and titration assays has been widely adopted for anti-drug antibody testing. GQ1001, a recombinant humanized anti-human epidermal growth factor receptor 2 monoclonal antibody covalently linked to a cytotoxin of DM1, possesses a novel format of antibody–drug conjugates. In this study, we reported the development, validation, and application of an acid-dissociation bridging enzyme-linked immunosorbent assay for the detection of antibodies against GQ1001 in cynomolgus monkey serum. The sensitivity of the screening assay was 126.141 ng/mL in undiluted serum. The screening assay and confirmatory assay were neither affected by the naïve monkey serum nor by 2% and 5% (v/v) erythrocyte hemolysates. Moreover, the assay was not subject to interference by 2500 ng/mL of human IgG1 in the samples. Drug interference at low positive control (150 ng/mL) and high positive control (8000 ng/mL) of anti-GQ1001 antibodies was not observed when GQ1001 concentrations were below 3.125 μg/mL and 100 μg/mL, respectively. Furthermore, no hook effect was observed for the positive antibodies in the concentration range of 8 to 64 μg/mL. The validated assay was, thereafter, successfully applied to a single-dose toxicity study of GQ1001. Anti-drug antibody positive rates among dosing animals and testing samples were reported, and no significant impact was found on toxicokinetic outcomes.
- Subjects
IMMUNOGLOBULINS; KRA; MONOCLONAL antibodies; EPIDERMAL growth factor receptors; ANTIBODY-drug conjugates; ENZYME-linked immunosorbent assay; ANTIBODY titer
- Publication
Molecules, 2023, Vol 28, Issue 4, p1684
- ISSN
1420-3049
- Publication type
Article
- DOI
10.3390/molecules28041684