抗口蹄疫病毒IgA抗体的构建表达及其抗病毒活性的分析

doi:10.11843/j.issn.0366-6964.2025.08.036

Abstract

Abstract:

Foot-and-mouth disease(FMD)caused by foot-and-mouth disease virus (FMDV) is a high infectious disease of pigs, cattle, sheep and other major livestock. In order to develop new strategies against FMDV to carry out comprehensive prevention and control of the disease, the heavy chain variable region (VH) sequences and light chain variable region (VL) sequences of 5 different IgA antibodies against type O and A FMDV were screened from the specific porcine BCR library of type O and A FMDV and linked with the gene sequences of the heavy chain constant region and the light chain (Kappa chain) constant region of porcine IgA, then, five heavy chain plasmids and five light chain plasmids were respectively constructed. The corresponding heavy chain plasmids and light chain plasmids were co-transfected into CHO-s suspension cells, after 7 days post transfection, the transfected supernatants were collected for antibody purification. The purified antibodies were identified by SDS-PAGE and Western blot. The ability of the five antibodies neutralizing types O and A FMDV was analyzed by virus neutralization test. The reactivity of the five antibodies with FMDV was detected by ELISA. Results showed that 5 IgA antibodies were successfully expressed in CHO-s cells and they all could not neutralize type O and A FMDV, but one of them (POA-A8) had the strongest ability to specifically bind with FMDV, which could be used for establishment of FMDV diagnostic methods in the future. This study lays a foundation for research of anti-FMDV nonneutralizing sIgA antibodies and development of diagnostic methods for FMDV in the future.

Key words: FMDV, IgA antibody, construction and expression, antiviral activity

CLC Number:

S852.659.6

LIU Feng, LI Kun, ZHANG Xingze, MA Xueqing, SUN Pu, LI Fengjuan, CAO Yimei, BAI Xingwen, FU Yuanfang, YUAN Hong, OUYANG Yifan, LIU Zaixin, LU Zengjun, LI Pinghua. Construction and Expression of IgA Antibodies against Foot-and-mouth Disease Virus and Analysis of Its Antiviral Activity[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3985-3991.

Figures/Tables 7

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References 15

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IgA重/轻链载体质粒 Heavy/ light chain plasmid of IgA	载体质粒名称 Vector plasmid	信号肽序列(aa) Signal peptide sequence	可变区编码长度/aa Code length of variable region
重链载体质粒 Heavy chain vector plasmid	POA-AH3	MEFRLHWVVLFALLQDVQG	143
	POA-AH5	MEFRLNWVVLFVLLQGVQG	138
	POA-AH6	MEFRLNWVVLFALLQGVQG	137
	POA-AH7	MEFRLNWVVLFALLQGVQG	132
	POA-AH8	MEFRLNWVVLFALLQGVQG	136
轻链载体质粒 Light chain vector plasmid	POA-AL3	MAWTVLLIGLLAVGSGVDS	127
	POA-AL5	MAWTVLLIGLLAVGSGVES	129
	POA-AL6	MAWTVLLIGLLAVGSGVDS	127
	POA-AL7	MAWTVLLIGLLAVGSGVDS	127
	POA-AL8	MAWTVLLIGLLAVGSGVDS	125

IgA抗体	IC₅₀
IgA抗体	O/BY/CHA/ 2010	O/HK99	O/GSLX	O/Mya/98	O/HN/ CHA/93	O/XJ/CHA/ 2017	A/WH/09	A/GDMM/ CHA/2013	A/AF72
PAO-A3	>50	>50	>50	>50	>50	>50	>50	>50	>50
POA-A5	>50	>50	>50	>50	>50	>50	>50	>50	>50
POA-A6	>50	>50	>50	>50	>50	>50	>50	>50	>50
POA-A7	>50	>50	>50	>50	>50	>50	>50	>50	>50
POA-A8	>50	>50	>50	>50	>50	>50	>50	>50	>50

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Construction and Expression of IgA Antibodies against Foot-and-mouth Disease Virus and Analysis of Its Antiviral Activity

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