Bioinformatics analysis of ureaplasma urealyticum UP3-RS02445 and the preparation of monoclonal antibodies

• Published in: Xi bao yu fen zi mian yi xue za zhi Vol. 40; no. 11; p. 1011
• Main Authors: Chen, Hengxin, Jia, Xiaohui, Li, Yahui, Zhou, Yan, Jia, Tianjun, Li, Ping
• Format: Journal Article
• Language: Chinese
• Published: China 01.11.2024
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Bacterial Proteins - genetics; Bacterial Proteins - immunology; Computational Biology - methods; Enzyme-Linked Immunosorbent Assay; Female; Hybridomas - immunology; Mice; Mice, Inbred BALB C; Ureaplasma urealyticum - genetics; Ureaplasma urealyticum - immunology
• ISSN: 1007-8738

Objective To make the bioinformatics analysis of Ureaplasma parvum UP3-RS02445 and prepare monoclonal antibody (mAb) against UP3-RS02445. Methods The biological characteristics of UP3-RS02445 protein were predicted by bioinformatics software. The UP3-RS02445 prokaryotic expression plasmid was constructed and the corresponding protein expression was induced by isopropyl-β-D-thiogalactoside (IPTG). Thus the expressed protein was used as immunogen to immunize female BALB/c mice. Hybridoma cell technology was used to prepare the monoclonal antibody against UP3-RS02445. The specificity and titer of monoclonal antibody were detected by Western blot and ELISA respectively. The subclass of heavy chain and subtype of light chain were identified by monoclonal antibody subtype identification test strip. Results Bioinformatics analysis showed that UP3-RS02445 protein was composed of 201 amino acids, without transmembrane domain and signal peptide, and belongs to non-secretory proteins. The recombinant prokaryotic plasmid