An Acid Dissociation Bridging ELISA Assay for Detection of Antibodies Against Therapeutic Monoclonal Antibody ArzerraTM in Cynomolgus Serum
WANG Bian-zhen1,2, DENG Cheng-lian2, ZOU Jia1, GAO Liu-cun1, DONG Bin2,SONG Hai-feng1*
1. Research center of Pharmacokinetics, Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China; 2. Beijing United-Power Pharma Tech Co., Ltd., Beijing 102206, China
Abstract��OBJECTIVE To develop and validate an acid dissociated bridging ELISA assay for determination of antibodies (ADAs) against a human monoclonal antibody against CD20, ArzerraTM, in cynomulgus serum, which will be used to reflect the immunogenicity of ArzerraTM in cynomolgus serum. METHODS Biotin labeled ArzerraTM and acid treated unknown samples were added into the ArzerraTM pre-coated 96-well plates, sequentially. Positive controls (ADAs) in unknown samples bridged with coated ArzerraTM and biotin-labeled ArzerraTM (Biotin-Drug) to generate ADAs bridged compounds (Drug-ADAs-Biotin-Drug), and coloration was made with enzymatic reaction. RESULTS The sensitivity was 7.4 ng��mL-1, and the intra-assay and the inter-assay RSDs were less than 17.6% for positive controls. The screening cut point (SCP) for this assay was 1.23 and the confirmatory cut point (CCP) was 42.8%. The assay also showed high drug tolerance, which was 20 fold at 2.0 ��g��mL-1 ADA level (Drug:ADA=40.0 ��g��mL-1:2.0 ��g��mL-1); and no ��Hook effect�� was detected within 3.2-100 000.0 ng��mL-1. Using this assay to determine ADAs against ArzerraTM in Cynomolgus serum, ADAs were detected at 21 d post administration. CONCLUSION The bridging ELISA is one of the best assays to determine ADAs against therapeutic monoclonal antibodies, because it shows high sensitivity and good drug tolerance.
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WANG Bian-zhen, DENG Cheng-lian, ZOU Jia, GAO Liu-cun, DONG Bin,SONG Hai-feng. An Acid Dissociation Bridging ELISA Assay for Detection of Antibodies Against Therapeutic Monoclonal Antibody ArzerraTM in Cynomolgus Serum. Chinese Pharmaceutical Journal, 2017, 52(1): 14-19.
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2017, Vol. 52 
