title: Soluble production and function of vascular endothelial growth factor/basic fibroblast growth factor complex peptide.
abstract: Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are important proangiogenic factors in tumor procession. The autocrine and paracrine bFGF and the VEGF in tumor tissue can promote tumor angiogenesis, tumor growth, and metastasis. A VEGF/bFGF Complex Peptide (VBP3) was designed on the basis of epitope peptides from both VEGF and bFGF to elicit in vivo production of anti-bFGF and anti-VEGF antibodies. In this study, we reported on the production of recombinant VBP3 using high cell density fermentation. Fed-batch fermentation for recombinant VBP3 production was conducted, and the production procedure was optimized in a 10-L fermentor. The fraction of soluble VBP3 protein obtained reached 78% of total recombinant protein output under fed-batch fermentation. Purified recombinant VBP3 could inhibit tumor cell proliferation in vitro and stimulate C57BL/6 mice to produce high titer anti-VEGF and anti-bFGF antibodies in vivo. A melanoma-grafted mouse model and an immunohistochemistry assay showed that tumor growth and tumor angiogenesis were significantly inhibited in VBP3-vaccinated mice. These results demonstrated that soluble recombinant VBP3 could be produced by large-scale fermentation, and the product, with good immunogenicity, elicited production of high-titer anti-bFGF and anti-VEGF antibodies, which could be used as a therapeutic tumor vaccine to inhibit tumor angiogenesis and tumor growth.
authors.affiliation: The State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhang
authors.forename: Xuejun
authors.initials: X
authors.lastname: Lao
authors.forename: Jianhua
authors.initials: J
authors.lastname: Huang
authors.forename: Zhongsong
authors.initials: Z
authors.lastname: Zhu
authors.forename: Lei
authors.initials: L
authors.lastname: Pang
authors.forename: Yong
authors.initials: Y
authors.lastname: Tang
authors.forename: Qifang
authors.initials: Q
authors.lastname: Song
authors.forename: Jiangfang
authors.initials: J
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Deng
authors.forename: Ning
authors.initials: N
authors.lastname: Deng
authors.forename: Qin
authors.initials: Q
authors.lastname: Yang
authors.forename: Aditi M
authors.initials: AM
authors.lastname: Sengupta
authors.forename: Likuan
authors.initials: L
authors.lastname: Xiong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019496:Cancer Vaccines / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D005285:Fermentation; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic / Q000473:pathology; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D019496:Cancer Vaccines; D010446:Peptide Fragments; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2
keywords: VEGF; bFGF; fermentation; polypeptide; proangiogenic factor; tumor; vaccination