Development, validation, and application of ELISA for detection of anti-HD105 antibodies in pre-clinical safety evaluation using monkeys.
Unwanted immunogenicity of protein therapeutics can result in severe side effects and should be assessed in animals before applying the treatment to humans. Monkeys are the most relevant choice for pre-clinical toxicity testing of antibody-based therapeutics. To assess the immunogenicity of HD105, a novel antibody therapeutic that targets both vascular endothelial growth factor and Delta-like-ligand 4, a bridging enzyme-linked immunosorbent assay was developed as an anti-drug antibody (ADA) assay and validated for use in pre-clinical studies using non-human primates. This method was found to have suitable assay sensitivity, intra- and inter-assay precision, confirmation, drug tolerance, recovery, and sample stability for measuring ADA in monkey serum samples. The results showed that ADA elevation occurred following repeated doses of HD105, and that ADA production was negatively associated with serum HD105 concentration. These results suggest that intravenous administration of HD105 induces production of ADA in monkeys and that the detection of ADA may be negatively influenced by free HD105 in serum.
College of Pharmacy, Chungnam National University, Daejeon, Republic of Korea; Korea Institute of Toxicology, 141 Gajeong-ro, Yuseong-gu, Daejeon, Republic of Korea.
Woo Hyuck
WH
Choi
Korea Institute of Toxicology, 141 Gajeong-ro, Yuseong-gu, Daejeon, Republic of Korea.
Hye Ryun
HR
Jo
Korea Institute of Toxicology, 141 Gajeong-ro, Yuseong-gu, Daejeon, Republic of Korea.
Eun-Jeong
EJ
Jeon
Korea Institute of Toxicology, 141 Gajeong-ro, Yuseong-gu, Daejeon, Republic of Korea.
So-Young
SY
Youm
College of Pharmacy, Chungnam National University, Daejeon, Republic of Korea.
Jang Su
JS
Jeon
Hanwha Chemical R&D Center, Biologics Business Unit, Gajeong-Ro, Yuseong-Gu, Daejeon, Republic of Korea.
Yong-Gyu
YG
Son
Hanwha Chemical R&D Center, Biologics Business Unit, Gajeong-Ro, Yuseong-Gu, Daejeon, Republic of Korea.
Weon-Kyoo
WK
You
Hanwha Chemical R&D Center, Biologics Business Unit, Gajeong-Ro, Yuseong-Gu, Daejeon, Republic of Korea.
Woo Suk
WS
Koh
Hanwha Chemical R&D Center, Biologics Business Unit, Gajeong-Ro, Yuseong-Gu, Daejeon, Republic of Korea.
Sang Hoon
SH
Lee
College of Pharmacy, Chungnam National University, Daejeon, Republic of Korea. Electronic address: sangkim@cnu.ac.kr.
Sang Kyum
SK
Kim
D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood* / Q000633:toxicity; D001323:Autoantibodies / Q000097:blood* / Q000187:drug effects; D002626:Chemistry, Pharmaceutical / Q000379:methods / Q000592:standards*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005260:Female; D006801:Humans; D008252:Macaca fascicularis; D008297:Male; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D001323:Autoantibodies
Anti-drug antibody; Bispecific antibody; Immunogenicity; Preclinical study; Therapeutic proteins; Validation