title: Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system.
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.
authors.affiliation: 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.
authors.forename: Bo
authors.initials: B
authors.lastname: Lin
authors.affiliation: 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.
authors.forename: Kun
authors.initials: K
authors.lastname: Liu
authors.affiliation: Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China.|Department of Anesthesiology, Second Affiliated Hospital, Hainan Medical College, Haikou 570311, PR China.
authors.forename: Wenting
authors.initials: W
authors.lastname: Wang
authors.affiliation: 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.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: 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.
authors.forename: Xu
authors.initials: X
authors.lastname: Dong
authors.affiliation: 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.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.affiliation: 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.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.affiliation: 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.
authors.forename: Junli
authors.initials: J
authors.lastname: Guo
authors.affiliation: Hainan Provincial Key Laboratory of Carcinogenesis and Intervention, Hainan Medical College, Haikou 571199, Hainan Province, PR China mingyuezhu2002@163.com mengsenli@163.com.|Key Laboratory of Molecular Biology, Hainan Medical College, Haikou 571199, PR China.
authors.forename: Mingyue
authors.initials: M
authors.lastname: Zhu
authors.affiliation: 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.|Institution of Tumor, Hainan Medical College, Haikou 570102, Hainan Province, PR China.
authors.forename: Mengsen
authors.initials: M
authors.lastname: Li
mesh_terms: D000818:Animals; D016367:Baculoviridae; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D006801:Humans; D007155:Immunologic Factors / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D007313:Insecta / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: C097353:AFP protein, human; D003470:Culture Media; D007155:Immunologic Factors; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins
keywords: Alpha fetoprotein; Bioactivity; Expressed vectors; Hepatoma cells