Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system

Author:

Lin Bo12,Liu Kun12,Wang Wenting13,Li Wei12,Dong Xu12,Chen Yi12,Lu Yan12,Guo Junli12,Zhu Mingyue12,Li Mengsen124

Affiliation:

1. Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China

2. Key Laboratory of Molecular Biology, Hainan Medical College, Haikou 571199, PR China

3. Department of Anesthesiology, Second Affiliated Hospital, Hainan Medical College, Haikou 570311, PR China

4. Institution of Tumor, Hainan Medical College, Haikou 570102, Hainan Province, PR China

Abstract

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.

Publisher

Portland Press Ltd.

Subject

Cell Biology,Molecular Biology,Biochemistry,Biophysics

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