重组人源化抗TNF-α单抗WLB303猴重复给药毒性试验血清中和抗体确证方法的建立

发布时间：2018-10-30 18:42

【摘要】：目的:建立一种快速、灵敏、高效和易操作的方法,用于检测重组人源化抗TNF-α单抗WLB303猴重复给药毒性试验血清中产生的中和抗体。方法:对食蟹猴静脉输注WLB303,连续给药4周。采取给药期和恢复期不同时间的血清,随后采用桥式酶联免疫吸附法对血清中的抗药抗体进行定性筛选,对筛选为阳性的血清样本进行预处理,然后以WLB303生物学活性测定法为基础的中和反应法对中和抗体进行确证。结果:建立了WLB303中和抗体的确证方法,确定了检测的临界点。确证方法的检出率为100%,在血清中检测到具有中和活性的抗体,中和抗体的中和作用强度总体与给药时间、给药剂量呈正相关。结论:建立的检测方法能够直接体现出剂量差异性和个体差异性,也能反映中和抗体的滴度差别。该方法的特异性、灵敏度和有效性能够满足试验要求,可以用于单抗药物重复给药动物毒性试验血清中和抗体的检测。
[Abstract]:Objective: to establish a rapid, sensitive, efficient and easy to operate method for the detection of neutralizing antibodies produced in the sera of recombinant human anti TNF- 伪 monoclonal antibody WLB303 monkeys. Methods: WLB303, was infused intravenously for 4 weeks. Serum samples of different periods of administration and convalescence were used, and then the anti-drug antibodies in serum were screened qualitatively by bridge enzyme-linked immunosorbent assay (Elisa), and the positive serum samples were pretreated. Then the neutralization reaction method based on WLB303 assay was used to confirm the neutralization antibody. Results: the method of WLB303 neutralizing antibody was established and the critical point of detection was determined. The positive rate of the method was 100. The neutralizing activity of antibody was detected in serum. The neutralization intensity of neutralizing antibody was positively correlated with the administration time and dosage. Conclusion: the established method can not only reflect the difference of dose and individual, but also the titer of neutralizing antibody. The specificity, sensitivity and effectiveness of the method can meet the requirements of the test, and can be used for the detection of serum neutralization antibody in repeated drug administration animal toxicity test.
【作者单位】：深圳万乐药业有限公司;北京昭衍新药研究中心股份有限公司;
【分类号】：R965

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