Lin Bo, Liu Kun, Wang Wenting, Li Wei, Dong Xu, Chen Yi, Lu Yan, Guo Junli, Zhu Mingyue, Li Mengsen
Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China.
Key Laboratory of Molecular Biology, Hainan Medical College, Haikou 571199, PR China.
Biosci Rep. 2017 Jan 17;37(1). doi: 10.1042/BSR20160161. Print 2017 Feb 28.
α-fetoprotein (AFP) is an early serum growth factor in foetal embryonic development and hepatic oncogenesis. A growing number of investigations of AFP as a tumour-specific biomarker have concluded that AFP is an important target for cancer treatment. AFP also plays an immunomodulatory role in the treatment of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, myasthenia gravis and thyroiditis. In an effort to support biochemical screening and drug design and discovery, we attempted to express and purify human AFP in a Bac-to-Bac system. Two key factors affecting the expression of recombinant human AFP (R-AFP), namely the infectious baculovirus inoculum volume and the culturing time post-infection, were optimized to maximize the yield. We achieved a high yield of approximately 1.5 mg/l of harvested medium with a 72-96 h incubation period after infection and an inoculum volume ratio of 1:100. We also assessed the role of R-AFP in the proliferation of the human liver cancer cell line Bel 7402, and the results indicated that R-AFP promoted the growth of hepatoma cells. We concluded that this method can produce high yields of R-AFP, which can be used for studies related to AFP.
甲胎蛋白(AFP)是胎儿胚胎发育和肝脏肿瘤发生过程中的一种早期血清生长因子。越来越多将AFP作为肿瘤特异性生物标志物的研究得出结论,AFP是癌症治疗的一个重要靶点。AFP在治疗几种自身免疫性疾病(如类风湿性关节炎、多发性硬化症、重症肌无力和甲状腺炎)中也发挥着免疫调节作用。为了支持生化筛查以及药物设计与发现,我们尝试在杆状病毒表达系统中表达并纯化人AFP。对影响重组人AFP(R-AFP)表达的两个关键因素,即感染性杆状病毒接种量和感染后培养时间进行了优化,以实现产量最大化。感染后经过72 - 96小时的孵育期以及接种量比例为一比一百的条件下,我们在收获的培养基中实现了约1.5毫克/升的高产量。我们还评估了R-AFP在人肝癌细胞系Bel 7402增殖中的作用,结果表明R-AFP促进了肝癌细胞的生长。我们得出结论,该方法能够高产R-AFP,可用于与AFP相关的研究。