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目的:通过序列分析确定抗胃癌抗体重链可变区(V_H)的一级结构,并借助同源模建方法模拟其三级结构.方法:从抗人胃癌噬菌体抗体库中筛选出V_H基因,并进行序列测定、翻译和分析.利用计算机辅助蛋白质空间模拟技术,采用同源模建、力学优化合理模建V_H的三维空间结构.结果:序列比对分析表明获得的V_H序列符合鼠抗体可变区特征,通过Kabat分析确定了FR、CDR;合理搭建了抗体重链可变区的空间构象,并通过分子力学优化获得了稳定的三维结构.结论:所测得的V_H一级结构和构建的三维空间结构均有较高的可靠性,为进一步的生物学实验奠定了基础.
OBJECTIVE: To determine the primary structure of heavy chain variable region (V_H) of anti-gastric cancer antibody by sequence analysis and to simulate its tertiary structure by means of homology modeling.Methods: V_H gene was screened from anti-human gastric phage antibody library, The three-dimensional structure of V_H was modeled by homology modeling and mechanics optimization.Results: Sequence alignment analysis showed that the obtained V_H sequence was in accordance with the variable of mouse antibody Region characteristics, the FR and CDR were determined by Kabat analysis, and the spatial conformation of antibody heavy chain variable region was reasonably established and the stable three-dimensional structure was obtained through molecular mechanics optimization.Conclusion: The measured V_H primary structure and the constructed The three-dimensional structure has higher reliability, which lays the foundation for further biological experiments.