title: Structural characterization of a recombinant fusion protein by instrumental analysis and molecular modeling.
abstract: Conbercept is a genetically engineered homodimeric protein for the treatment of wet age-related macular degeneration (wet AMD) that functions by blocking VEGF-family proteins. Its huge, highly variable architecture makes characterization and development of a functional assay difficult. In this study, the primary structure, number of disulfide linkages and glycosylation state of conbercept were characterized by high-performance liquid chromatography, mass spectrometry, and capillary electrophoresis. Molecular modeling was then applied to obtain the spatial structural model of the conbercept-VEGF-A complex, and to study its inter-atomic interactions and dynamic behavior. This work was incorporated into a platform useful for studying the structure of conbercept and its ligand binding functions.
authors.affiliation: Chengdu Kanghong Biotechnology Inc, Chengdu, PR China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wu
authors.forename: Peng
authors.initials: P
authors.lastname: Zhou
authors.forename: Xiaoxin
authors.initials: X
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Delun
authors.initials: D
authors.lastname: Luo
authors.forename: Huaiyao
authors.initials: H
authors.lastname: Qiao
authors.forename: Xiao
authors.initials: X
authors.lastname: Ke
authors.forename: Jian
authors.initials: J
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D018360:Crystallography, X-Ray; D019075:Electrophoresis, Capillary; D006031:Glycosylation; D013058:Mass Spectrometry; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D011034:Podophyllotoxin / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors / Q000737:chemistry*
chemical_list: D011993:Recombinant Fusion Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D011034:Podophyllotoxin