State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, 100193, China.
Transgenic Res. 2012 Oct;21(5):1085-91. doi: 10.1007/s11248-012-9589-z.
Anti-HBV monoclonal antibodies (mAbs) have been successfully applied in clinic for the prevention and treatment of liver diseases caused by Hepatitis B virus (HBV). However, the mammalian cell culture technique used to produce mAbs provides low yield. Production capacity may be increased by manipulating the mammary gland of transgenic animals to produce mAbs which may be secreted in the milk and readily purified. In this study, we established a transgenic mouse model expressing anti-HBV mAbs and assessed the quantity and quality of such in milk secretions. Eight transgenic founders were generated by co-microinjection of two gene cassettes encoding the heavy- and light-chain of a neutralizing anti-HBV antibody. Both the heavy- and light-chain transgenes produced sufficient transcript expression in the transgenic mice. The expressed anti-HBV mAb was correctly assembled and modified in the mammary gland, as detected by western blotting. ELISA was used to determine the expression level of anti-HBV mAb, which reached up to 17.8 mg/mL, and the affinity of anti-HBV mAb for HBV surface antigen HBsAg, which reached up to 1,577 IU/mg. Therefore, our work confirms the concept of using transgenic animals for large-scale production of therapeutic anti-HBV mAb.
抗乙型肝炎病毒（HBV）单克隆抗体（mAbs）已成功应用于临床，用于预防和治疗乙型肝炎病毒（HBV）引起的肝脏疾病。然而，用于生产 mAbs 的哺乳动物细胞培养技术产量较低。通过操纵转基因动物的乳腺来生产 mAbs，可以增加生产能力，mAbs 可以在乳汁中分泌，并易于纯化。在本研究中，我们建立了表达抗 HBV mAbs 的转基因小鼠模型，并评估了乳汁中此类 mAbs 的产量和质量。通过共显微注射两个基因盒，生成了 8 个转基因 Founder，这两个基因盒编码中和抗 HBV 抗体的重链和轻链。重链和轻链转基因在转基因小鼠中均产生了足够的转录表达。Western blot 检测到 mAb 在乳腺中正确组装和修饰。ELISA 用于测定抗 HBV mAb 的表达水平，高达 17.8mg/mL，以及抗 HBV mAb 与乙型肝炎表面抗原 HBsAg 的亲和力，高达 1577IU/mg。因此，我们的工作证实了使用转基因动物大规模生产治疗性抗 HBV mAb 的概念。
