A CHO cell line expressing PEDV GIIb isoform S protein exocrinely and its application

By employing a codon-optimized TPA signal peptide and chimeric signal peptide strategy in the CHO cell system, a stable exocrine expression system for PEDV GIIb S protein was established, solving the problems of low expression efficiency and poor stability in the CHO cell system. This achieved efficient and stable PEDV GIIb S protein expression, meeting the needs of large-scale vaccine production.

CN122128243APending Publication Date: 2026-06-02NINGXIA UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NINGXIA UNIVERSITY
Filing Date
2026-03-17
Publication Date
2026-06-02

AI Technical Summary

Technical Problem

Existing technologies for expressing PEDV GIIb subtype S protein in CHO cell systems suffer from low expression efficiency, poor stability, and insufficient capacity for large-scale production. In particular, during exocrine expression, the binding capacity of conventional signal peptides and the efficiency of endoplasmic reticulum transport are limited, leading to problems such as high protein retention, increased purification difficulty, and high production costs.

Method used

The codon-optimized TPA signal peptide guides the exocrine expression of PEDV GIIb S protein. By constructing a recombinant vector containing a chimeric signal peptide, the PEDV GIIb S protein encoding gene, and a dual-selection marker, CHO-S cells were transfected and antibiotics were screened to establish a stable chromosome-integrated expression system. This ensured that the protein was expressed in a secretory form and constructed a stable trimer structure. The signal peptide selection strategy was optimized to improve secretion efficiency.

Benefits of technology

It significantly improved the exocrine expression level of PEDV GIIb S protein, ensuring stable high expression in CHO cell lines during multiple generations of culture, reducing production costs and improving purification convenience, thus meeting the standardization and large-scale requirements of vaccine production.

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Abstract

This invention relates to the field of vaccine technology and provides a CHO cell line for exocrine expression of PEDV GIIb subtype S protein and its application. The CHO cell line integrates a codon-optimized PEDV GIIb S protein encoding gene carrying the TPA signal peptide into its chromosome. The nucleotide sequence of the PEDV GIIb S protein encoding gene is shown in SEQ ID NO.1. This invention uses currently prevalent GIIb strains as the antigen source to ensure vaccine strain matching; it constructs a secretory expression structure, thereby significantly improving the exocrine expression efficiency of the S protein; it precisely truncates the transmembrane region while retaining key structural domains, maintaining the protein's native conformation and immunogenicity; and it obtains a CHO cell line with stable gene integration and long-term passage to maintain expression levels through optimized transfection conditions and an antibiotic selection system.
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