Study on the Relationship between the Structure and Functions of Antihuman Cervical Cancer Single-chain Antibody and the Lengths of Linkers

Abstract

IntroductionCervical cancer (CC) is a malignant tumor occurred in the uterus vagina and cervical canals. It is one of the most common gynecological malignancies [1] and the incidence takes the second place among gynecologic oncology. Each year, there are about 530,000 new cervical cancer cases worldwide, of which 85% occur in developing countries, and cervical cancer is one of the main causes of death of women in developing countries [2][3][4]. Therefore, how to improve the long-term survival of patients with cervical cancer is an important issue in clinical and basic research.With constantly thorough study of molecular biology, genetically engineered antibody has been widely used in the diagnoses and treatments of diseases. It has great potential in the diagnosis and treatment of malignancy. Single-chain variable fragment (ScFv) is a small molecule antibody that has intact antigen-binding site. Compared with the defects of monoclonal antibody, such as large amount of antibody molecules, low penetrating ability, low clearance rate of blood and low ration of tumor blood [5], ScFv has advantages, such as small amount of antibody molecules, high penetrating ability, low immunogenicity and easy to mass production. It can also neutralize virus and toxin to give play to immunization in cells and be guided carrier. However, studies about the application of ScFv in CC treatment are rarely reported. As a result, it is of particular significance to conduct an in-depth research on the single-chain antibody of CC, which would provide a new direction of treating CC.ScFv is recombinant protein linked by a length of connecting peptide through VH and VL. Different lengths of linkers among chains have different influence on the active of single-chain antibody, so an insight into the impact of different lengths of linkers among chains on the biological activity of single-chain antibody has great significance in the establishment of single-chain antibody and screening of linkers among chains. By using the bioinformatics method, this study compared the impact of different linkers among chains on the structures and functions of single-chain antibody of CC from the aspects of protein tertiary structure, aiming to providing new ideas of the screening of interchain linker length of CC and the establishment of single-chain antibody, and also providing some theoretical bases for the targeted therapy of CC. Materials and Methods Experimental materialsThe amino acid sequences of single-chain antibody VH and VL of CC were from the articles related to researches of the anti-human cervical cancer written by Wang et al. [6]. Hela human cervical epithelial cell line was preserved in our lab. We ourselves constructed the general expression vector of single-chain antibody of connecting peptide with different length, expressed by HB2151 expressing system, AbstractBackground: This study aimed to find the linker length with minimal impact among chains to fight against the structure and function of cervical cancer single-chain antibody.