Comparison of two ELISA methods for detecting anti-drug antibody level against anti-tetanus toxin monoclonal antibodies in rhesus monkeys
10.13200/j.cnki.cjb.004444
- VernacularTitle:两种ELISA法检测恒河猴体内针对抗破伤风毒素单克隆抗体的抗药物抗体水平的比较
- Author:
XING Shanshan
- Publication Type:Journal Article
- Keywords:
Monoclonal antibodies against tetanus toxin;
Anti-drug antibody(ADA);
Bridging ELISA;
Affinity capture elution ELISA(ACE ELISA)
- From:
Chinese Journal of Biologicals
2025;38(03):359-356
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect the level of anti-drug antibodies(AD As) against anti-tetanus toxin monoclonal antibodies in rhesus monkeys by bridging ELISA and affinity capture elution ELISA(ACE ELISA),and compare them in order to determine an efficient and rapid method for the detection of ADA.Methods The ADA level of anti-tetanus toxin monoclonal antibodies was detected by bridging ELISA and ACE ELISA,and the serum minimum dilution multiple,screening cut point(SCP) and confirmation cut point(CCP) were determined.The sensitivity,drug resistance and precision of the two methods were verified.Forty rhesus monkeys(half male and half female) were randomly divided into negative control group,low dose group,high dose group and intravenous group.The negative control group was inoculated with normal saline,while the other groups were given different doses of anti-tetanus toxin monoclonal antibodies.The blood samples were collected from limbs veins before administration,14 and 28 d after the initial administration and at the end of the recovery period,and the serum was separated,which was detected for the ADA by two methods.Results The minimum dilution multiple of serum was determined to be 1:10.The SCPs of bridging ELISA and ACE ELISA were 0.059 3 and 0.098 1,and the CCPs were 14% and 30%,respectively.The sensitivity of both bridging ELISA and ACE ELISA was 500 ng/mL,but the drug resistance of ACE ELISA at high concentration of positive control antibody(300 μg/mL) was better than that of bridging ELISA(480 ng/mL).The screening precision of bridging ELISA(<5%) was better than that of ACE ELISA(12.35%-21.36%).Four positive samples were detected in rhesus monkey serum samples by ACE ELISA,and three positive samples were confirmed;Eight positive samples were screened by bridging ELISA,while only one was positive after confirmation.The false positive rate of ACE ELISA(0.7%)was lower than that of bridging ELISA(5.2%).Conclusion ACE ELISA has high drug resistance in detecting anti-tetanus toxin monoclonal antibody ADA,which is superior to bridging ELISA,especially in the case of high drug concentration.Although ACE ELISA has slightly lower precision,it is still a reliable ADA detection method,suitable for preclinical research and clinical application of anti-tetanus toxin monoclonal antibodies
