High density fermentation and immune evaluation of porcine circovirus type 3 Cap protein
10.16303/j.cnki.1005-4545.2024.12.01
- VernacularTitle:猪圆环病毒3型Cap蛋白的高密度发酵工艺及免疫评价
- Author:
Kele LI
1
;
Jinglong XU
1
;
Xiaolin GENG
1
;
Zhifei WANG
1
;
Yanyu LIANG
1
;
Xiaolin MO
1
;
Yuxin HUANG
1
;
Wenqiang PANG
1
;
Kegong TIAN
1
Author Information
1. 国家兽用药品工程技术研究中心,河南洛阳 471003;普莱柯生物工程股份有限公司,河南洛阳 471003
- Publication Type:Journal Article
- Keywords:
PCV3 Cap protein;
acetic acid;
combined control strategy of DO and OUR;
high density fermentation
- From:
Chinese Journal of Veterinary Science
2024;44(12):2507-2513
- CountryChina
- Language:Chinese
-
Abstract:
In order to achieve high-efficiency expression of the porcine circovirus type 3 Cap protein in recombinant Escherichia coli,dissolved oxygen(DO)control strategy,oxygen uptake rate(OUR)control strategy and combined control strategy of DO and OUR were selected to investi-gate their effect on the expression of target protein in a 10 L fermenter.The high-density fermenta-tion process of recombinant E.coli was determined by investigating the best control range of OUR.The recombinant protein was identified by SDS-PAGE and Western blot,and the immunoge-nicity of the protein was evaluated by the animal experiment.The results showed that when the DO control strategy and OUR control strategy were used,the expression of target protein was low.Acetic acid was the key factor affecting the expression of target protein.When acetic acid concen-tration reached 1.02 g/L,Cap protein yield was reduced by 55.8％.In order to reduce the produc-tion of acetic acid,DO was selected to control oxygen supply before feeding,and OUR was selected as the parameter to control oxygen supply after feeding.When OUR value was maintained at 140-160 mmol/(L·h),the Cap protein yield was up to 650 mg/L.Western blot analysis confirmed that PCV3 Cap possessed antigenicity and specificity.Animal experiment showed that the antibody pos-itive rate was 100％after the second immunization,indicating that the target protein had better immunogenicity.The high density fermentation controlled by the combined control strategy of DO and OUR could achieve efficiently soluble expression of PCV3 Cap in the fermenter,which laid the foundation for the vaccine development.
