Abstract

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_s1-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.

抽象的

Nivolumab是一种完整的人IgG4抗程序性细胞死亡1（PD-1）抗体，最近成为临床上最受欢迎和成功的治疗性单克隆抗体之一。随着对Nivolumab和其他治疗性单克隆抗体的需求增加，除哺乳动物细胞培养外，乳腺生物反应器已被视为生产重组单克隆抗体的另一选择。在这里，我们在转基因小鼠的乳腺中表达了重组人抗PD-1抗体。构建了两个表达载体，分别在牛_αs1的控制下分别带有抗PD-1抗体的重链和轻链。-酪蛋白启动子。然后通过共显微注射两个表达盒产生转基因小鼠。获得了三个具有重链和轻链阳性的F0创建者。检测到两条链的转基因均稳定地传递给后代。在转基因小鼠的牛奶中检测到重组抗体，其最高表达水平高达80.52±0.82 mg / L，并且可以与人PD-1抗原特异性结合。因此，我们的结果表明在转基因动物的乳汁中生产抗PD-1抗体的可行性。