State Key Laboratory of Pathogens and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, 100071 Beijing, People's Republic of China.
Cancer Immunol Immunother. 2010 Feb;59(2):239-46. doi: 10.1007/s00262-009-0744-1.
A new VEGF receptor fusion protein FP3 was shown to have promising antitumor potency better than Bevacizumab. Characterization of its immune response is essential to the safe and effective administration in clinical trials. In this study, both BIACORE and ELISA assays were employed to assess pre-clinical immunogenicity of FP3 in monkeys.
Serum samples from 20 rhesus monkeys were analyzed for the generation of anti-FP3 antibody after intravenous administration of three doses of FP3 (n = 6 per group) or buffer control (n = 2). Sera samples were obtained at 2, 4, 6, 8, 10 weeks after the first administration.
It showed BIACORE presented linear correlation with the dilution of anti-FP3 antibody and the results of ELISA. Two weeks after the initial FP3 injection, anti-FP3 antibody was detected in about 20% FP3-treated monkeys. The ratio of positive samples and the titer of antibody increased along with the FP3-treatment time. Six weeks following FP3 injection almost all the samples were anti-FP3 antibody positive. Moreover, the titer of anti-FP3 antibody but not the ratio of positive samples was also enhanced when the dose of FP3 was elevated. Furthermore, the immunoglobulin types and subclasses of anti-FP3 antibody serum components were mainly identified as IgG1 and IgG4, not IgM. Serum antibodies are characterized that they could not block FP3 binding to VEGF and were non-neutralizing.
Our data implied that proteins with full human sequences may also have the potential to induce immune response in rhesus monkeys, and BIACORE could be an effective approach to detect the immunogenicity of protein therapeutics in clinic.
一种新的 VEGF 受体融合蛋白 FP3 显示出比 Bevacizumab 更好的抗肿瘤活性。其免疫反应的特征对于临床试验中的安全有效给药至关重要。在这项研究中，使用 BIACORE 和 ELISA 测定法来评估 FP3 在猴子中的临床前免疫原性。
分析了 20 只恒河猴静脉注射三剂 FP3（每组 6 只）或缓冲液对照（每组 2 只）后产生抗 FP3 抗体的血清样本。在首次给药后 2、4、6、8、10 周时获得血清样本。
BIACORE 与抗 FP3 抗体的稀释度呈线性相关，与 ELISA 的结果一致。在初始 FP3 注射后两周，约 20%的 FP3 治疗猴子中检测到抗 FP3 抗体。阳性样本的比例和抗体滴度随着 FP3 治疗时间的增加而增加。在 FP3 注射 6 周后，几乎所有的样本都是抗 FP3 抗体阳性。此外，当 FP3 剂量增加时，抗 FP3 抗体的滴度而非阳性样本的比例也增加。此外，抗 FP3 抗体血清成分的免疫球蛋白类型和亚类主要鉴定为 IgG1 和 IgG4，而不是 IgM。血清抗体的特点是它们不能阻断 FP3 与 VEGF 的结合，并且是非中和性的。
我们的数据表明，具有全长人序列的蛋白质也可能在恒河猴中具有诱导免疫反应的潜力，BIACORE 可能是一种有效的方法来检测蛋白质治疗药物在临床上的免疫原性。
