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Structural characterization of a recombinant fusion protein by instrumental analysis and molecular modeling.

PloS one (2013-03-08)
Zhigang Wu, Peng Zhou, Xiaoxin Li, Hui Wang, Delun Luo, Huaiyao Qiao, Xiao Ke, Jian Huang
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Podophyllotoxin
Sigma-Aldrich
Picropodophyllotoxin, ≥96% (HPLC), powder