China State Institute of Pharmaceutical Industry, Shanghai, P. R. China.
Prep Biochem Biotechnol. 2021;51(2):183-190. doi: 10.1080/10826068.2020.1805755. Epub 2020 Aug 18.
Nivolumab, a fully human IgG4 anti-programed cell death 1（PD-1）antibody, is recently one of the most popular and successful therapeutic monoclonal antibodies in clinical use. With the increasing demands for Nivolumab and other therapeutic monoclonal antibodies, the mammary gland bioreactor has been regarded as another choice for the production of recombinant monoclonal antibodies besides mammalian cell culture. Here, we expressed a recombinant human anti-PD-1 antibody in the mammary glands of transgenic mice. Two expression vectors were constructed bearing the heavy and light chains of anti-PD-1 antibody respectively under the control of bovine α-casein promoter. Transgenic mice were then generated by co-microinjection of the two expression cassettes. Three F0 founders with both heavy chain and light chain positive were obtained. Transgenes of both chains were detected to be stably transmitted to the offspring. The recombinant antibody was detected in the milk of transgenic mice with the highest expression level up to 80.52 ± 0.82 mg/L and could specifically binds to the human PD-1 antigen. Therefore, our results suggest the feasibility of anti-PD-1 antibody production in the milk of transgenic animals.
尼伏鲁单抗是一种全人源 IgG4 抗程序性细胞死亡蛋白 1（PD-1）抗体，是目前临床应用中最受欢迎和最成功的治疗性单克隆抗体之一。随着对尼伏鲁单抗和其他治疗性单克隆抗体需求的增加，乳腺生物反应器已被视为除哺乳动物细胞培养之外生产重组单克隆抗体的另一种选择。在这里，我们在转基因小鼠的乳腺中表达了一种重组人抗 PD-1 抗体。两个表达载体分别在牛α-酪蛋白启动子的控制下携带抗 PD-1 抗体的重链和轻链。然后通过共显微注射这两个表达盒来产生转基因小鼠。获得了 3 只同时带有重链和轻链阳性的 F0 创始鼠。检测到两条链的转基因都稳定地传递给了后代。在表达水平最高可达 80.52 ± 0.82mg/L 的转基因小鼠的乳汁中检测到重组抗体，并能特异性结合人 PD-1 抗原。因此，我们的结果表明在转基因动物的乳汁中生产抗 PD-1 抗体是可行的。
