稳定表达猪圆环病毒2型Cap蛋白的CHO-K1细胞系的建立及免疫原性分析

doi:10.11843/j.issn.0366-6964.2019.05.016

Abstract

Abstract:

In this study, we established a Chinese Hamster Ovary (CHO-K1) cell line stably expressing PCV2-Cap, and identified the immunogenicity of the protein and prepared for a kind of novel subunit vaccine against porcine circovirus. Firstly, we constructed the recombinant plasmid pEE12.4-PCV2-Cap. Then we transfected adherent CHO-K1 cells with the plasmid. To obtain the monoclonal cell line, the transfected cells were selected with the methods of pressurized screening and limited dilution. And then the monoclonal cell was optimized for growth in suspension and cultured in fed-batch mode for the quality evaluation. The cell strain was fermented, and the purified target protein was verified by immunoassay in mice. The results demonstrated that PCV2-Cap protein was correctly expressed by CHO-K1 cells. The maximum density of living cells reached 6×10⁶ per milliliter, the cell viability was more than 80% in the process of the culture, the PCV2-Cap production can get to 370 mg·L^-1. The results of ELISA showed that Cap protein produced by CHO-K1 has good immunogenicity. A CHO-K1 cell line stably expressing PCV2-Cap protein has been established, and the immunogenicity analysis of the target protein was further carried out. It lays a foundation for the development of new porcine circovirus subunit vaccine.

CLC Number:

S852.659.2

WU Leyi, WU Sufang, CHE Ying, ZHANG Qiang, LI Peng, BIAN Guanglin, WANG Xuannian. The Establishment of CHO-K1 Cell Line Stably Expressing High Level PCV2-Cap and Immunogenicity Analysis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1056-1063.

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The Establishment of CHO-K1 Cell Line Stably Expressing High Level PCV2-Cap and Immunogenicity Analysis

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