Expression and biological activity of recombinant long-acting feline IFNω-FSA fusion protein
10.16303/j.cnki.1005-4545.2025.10.12
- VernacularTitle:重组长效猫IFNω-FSA融合蛋白表达与生物活性
- Author:
Yanan ZHAO
1
;
Dandan YANG
1
;
Yudie ZHANG
1
;
Jinling GUO
1
;
Dongyu LIU
1
;
Yuhe YIN
1
;
Congmei WU
1
Author Information
1. 长春工业大学化学与生命科学学院,吉林长春 130012
- Publication Type:Journal Article
- Keywords:
feline serum albumin;
feline ω interferon;
fusion expression;
biological activity;
half-life
- From:
Chinese Journal of Veterinary Science
2025;45(10):2179-2186
- CountryChina
- Language:Chinese
-
Abstract:
A long-acting feline ω-interferon fusion protein(FSA-FeIFNω)was designed and its bio-logical function validated.According to the optimization of the sequence of feline serum albumin and feline ω interferon in NCBI,the recombinant plasmid pET-30a(+)-FSA-FeIFNω was con-structed,which was transformed into E.coli BL21(DE3)competent cells,the expression of re-combinant protein FSA-FeIFNω was induced by IPTG,and the expressed inclusion body protein was identified by Western blot,the refolding product was purified by Ni-NTA affinity chromatog-raphy,and the concentration of dialysis and concentrated protein after purification was determined by BCA method.The antiviral activity of recombinant protein was detected by micro-cytopathic in-hibition method in the CRFK/VSV system,the in vitro half-life was detected by 50％mouse plas-ma method,the tumor cell proliferation inhibition activity was detected by MTT method,and anti-tumor activity was detected by mouse melanoma model.The pET-30a(+)-FeIFNω and pET-30a(+)-FSA-FeIFNω expression vectors were successfully constructed,and 87 kDa recombinant FSA-FeIFNω protein was obtained in E.coli,with a purified protein purity of 95％,with a concen-tration of 1 g/L,and the biological activity was 2.56 × 106 IU/mg,the plasma half-life was significantly prolonged(＞24 h),and the half-inhibitory concentration IC50 of B16-F10 in mouse melanoma cells was 56.01 mg/L.The FSA-FeIFNω group significantly inhibited tumor growth,and the treatment effect was better than that of the control group and other experimental groups.The recombinant FSA-FeIFNω protein obtained in this study had long-acting effect and good biological activity.
