Effects of different signal peptides on the secretion of human-mouse chimeric CMV-IgM.
- Author:
Yamin CUI
1
;
Xiaoping TIAN
1
;
Qiaohui ZHAO
1
;
Guilin LI
1
Author Information
- Publication Type:Journal Article
- Keywords: CMV-IgM; expression; human-mouse chimeric; signal peptide
- MeSH: Animals; Antibodies, Viral; genetics; immunology; Cricetinae; Cytomegalovirus; immunology; Enzyme-Linked Immunosorbent Assay; Gene Expression; Humans; Immunoglobulin M; immunology; Mice; Protein Sorting Signals; Recombinant Fusion Proteins; immunology
- From: Chinese Journal of Biotechnology 2020;36(6):1223-1231
- CountryChina
- Language:Chinese
- Abstract: In order to prepare human-mouse chimeric cytomegalovirus-immunoglobulin M (CMV-IgM) in vitro and study the effects of different signal peptides on the secretion of CMV-IgM, genes were amplified from hybridoma cell line using RLM-RACE to construct the expression vector of chimeric CMV-IgM. Then, the signal peptide of SigF itself was replaced by five different secreted signal peptides (SigA-SigE) by PCR method, and the CHO cell was chosen as host cell for in vitro expression. SDS-PAGE, SEC-HPLC and ELISA experiments were carried out to evaluate the protein expression level and immunoreactivity of the purified CMV-IgM. A 910 kDa recombinant protein was successfully prepared and signal peptides (SigA-SigE) had an increased expressed CMV-IgM, which were 6.72, 5.19, 1.44, 1.85 and 1.98 times higher than that of the CMV 6# cell signal peptide SigF. In summary, this work provides a theoretical basis for the development of human-mouse chimeric CMV-IgM, and a novel route to increase the expression level of CMV-IgM.
