Development of a method for detection of serum neutralizing antibodies in cynomolgus monkeys undergoing repeated dose toxicity study of humanized anti-TNF-α monoclonal antibody WLB303
CHEN Hong-xia
LI Xue
LUO Tao
LI Le
YU Yu-gen
MENG Zhi-chao
LIU Si-si
Abstract：Objective:To establish a rapid,sensitive,highly-efficient and easy-to-use method for detection of neutralizing antibodies against WLB303 in serum of animals undergoing repeated dose toxicity study.Methods:The cynomolgus monkeys were continuously infused with recombinant humanized anti-TNF-α monoclonal antibodies intravenously for 4 weeks.Serum samples were taken at different time points before and after administration and at different stages of recovery period.Then a bridging-ELISA method was used to qualitatively screen anti-drug antibody (ADA).A sample pre-treatment procedure was conducted to eliminate the non-specific interference in serum samples prior to verification of neutralizing activity.Neutralization activity was verified in a cell-based neutralization antibody assay.Results:A cell-based neutralizing antibody assay was established and the cut-off point was confirmed.The detection rate of the confirmed neutralizing antibodies was 100％.Specific neutralizing antibodies were detected in serum samples,and the neutralization activity of neutralizing antibodies had positive correlation with the time and dosage of administration.Conclusion:The established assay has proven to be capable of demonstrating dosedependent effect difference and inter-individual difference.It also reflects difference in titers of neutralizing antibodies.The cell-based neutralizing antibody assay was found to have desired properties of specificity,sensitivity and validity,and can be used as a feasible method for detection of neutralizing antibodies in repeated dose toxicity study.
Keywords：bridging-ELISA methodneutralizing antibodybiological activityrepeated dose toxicity study
Publication Date：2017-01-01
Online Publishing Date：2026-08-14(First online date of this platform, not the publication date of the document)
Pages：10( 2386-2395 )