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Study on the Molecular Mechanism of Anti-CDK4 Single-chain Antibody

Author: WangMeiNiang
Tutor: LiGuiYing
School: Jilin University
Course: Biochemistry and Molecular Biology
Keywords: CDK4 Single-chain antibody Epitope Phage Peptide Library Computer Simulation
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 56
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Abstract


Studies have shown that malignant proliferation of tumor cells was mainly due to the imbalance caused by the cell cycle, Cyclin D-CDK4/CDK6 is the regulation of the cell cycle from G1 to S phase transition of the key factors. CDK4 gene has been shown to be an oncogene, in a variety of tumor cells and cancerous tissue are found accompanied by overexpression of CDK4 and excessive activation and, therefore, CDK4 gene therapy can be used as another potential target. Our group in the early work, have access to anti-CDK4 and single-chain antibody into tumor cells, successfully reversing the malignant proliferation of tumor cell phenotype, but we are for the single-chain antibody identify exactly the manner in which and the role of the antigen molecule that inhibits a target protein function, thereby exert its anti-tumor effect at the gene level mechanism problems still unclear. Epitope on the antigen represents an immune active region, the base is an antigen recognized by the antibody, with the biological techniques, epitope prediction for a variety of methods have been developed, in which its high-throughput screening of phage, easy to operate, it could also predict conformational epitopes and linear epitopes, etc., now has been widely used in various studies, especially on single-chain antibody phage libraries as a target protein to analyze its corresponding antigen epitope has become the current forecast antigen protein epitopes important ways. Because each epitope prediction method has some limitations, so using computational prediction (as source modeling, molecular docking, etc.) and biological method of combining experimental data antigen protein conformational epitope analysis and positioning has become the current research focus, and showing a trend of rapid growth. This study were prepared with biological activity and anti-CDK4 CDK4 protein antigens of human single-chain antibody (scFv) protein, thereby targeting with scFv from phage random peptide library screening ring seven to get a set of specific binding of anti- CDK4 in human single-chain antibody genes heptapeptide ring and ring with this group of biology software heptapeptide sequence alignment analysis performed. In order to obtain more accurate data support and a more intuitive way to understand protein interactions, we then homology modeling of the spatial structure of scFv with Builter software to draw a three-dimensional structure heptapeptide ring, and then use a computer docking methods simulate the interaction of small peptides and scFv spatial structure. According phage screening and computer simulation results, we hypothesized that small peptides screened by phage obtained in the analysis of epitopes epitopes part of CDK4 or activity of CDK4 in spatial conformation of epitopes formation plays a very important role.

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