猫杯状病毒VP1蛋白的单克隆抗体制备及抗原表位鉴定

doi:10.11843/j.issn.0366-6964.2024.12.040

Abstract

Abstract:

The purpose of this study was to prepare and identify monoclonal antibodies (mAbs) against the VP1 protein of feline calicivirus (FCV) strain SH14. In this study, the whole virus of the SH14 strain of FCV was used as an immunogen to immunize BALB/c mice through intraperitoneal injection, and mAbs against the VP1 protein were prepared. Through ELISA screening, a specific monoclonal antibody clone, 3A3C1, was successfully obtained. A comprehensive biological characterization of the 3A3C1 clone mAb was conducted, confirming it as an IgG₁ type with a κ light chain. The results of Western blot and immunofluorescence assay (IFA) showed that the 3A3C1 clone mAb could specifically recognize and bind to FCV strains SH14, F9, and GZ22. In addition, this study further explored the linear antigenic epitope of the 3A3C1 clone mAb. By segmentally expressing the VP1 protein and conducting detailed Western blot analysis, the linear antigenic epitope of the 3A3C1 clone mAb was finally determined to be located in the⁴²⁶PSRLTPAGDYAITSG⁴⁴⁰ region of the VP1 protein. The 3A3C1 clone mAb prepared in this study is not only an important tool for understanding the immunological characteristics of FCV but also has potential applications in developing FCV diagnostic methods and vaccine strategies.

Key words: feline calicivirus, monoclonal antibody, antigenic epitope, capsid protein

CLC Number:

S852.659.6

ZHANG Zeyu, DONG Ningning, TAN Xiaomei, LI Chuanfeng, ZHU Jie, LIU Guangqing, ZHANG Wei, MENG Chunchun. Preparation of Monoclonal Antibodies against Feline Calicivirus VP1 Protein and Identification of Antigenic Epitopes[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5784-5791.

Figures/Tables 6

Table 1

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References 20

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引物名称 Primers	序列(5′→3′) Sequences	产物大小/bp Product size
P1(426-547)	F: ctcggtaccctcgagggatccCCGAGCCGTCTGACCCCG R: agactgcaggtcgacaagcttACGAACCACTTTGTCACGATCC	366
P2(547-668)	F: ctcggtaccctcgagggatccATTAGCGTGCTGCCGGAAA R: agactgcaggtcgacaagcttAGCCCGGGATCCCAGTTT	366
P3(386-611)	F: ctcggtaccctcgagggatccACCACCGCGACCATCAGC R: agactgcaggtcgacaagcttGATACGATAAACCGCCAGGCTGTCCGGCGCCAGCAGGT	378
P4(426-456)	F: ctcggtaccctcgagggatccCCGAGCCGTCTGACCCCG R: agactgcaggtcgacaagcttGATAAAGCCGGTGTTGCTAACTT	93
P5(456-486)	F: ctcggtaccctcgagggatccACCACCGCGACCATCAGC R: agactgcaggtcgacaagcttACGAACCACTTTGTCACGATCC	93
P6(441-471)	F: ctcggtaccctcgagggatccATTAAGAACAACACCAACTTCAAAAGC R: agactgcaggtcgacaagcttTTTGCTCACACGGTTGTCTTGG	93

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Preparation of Monoclonal Antibodies against Feline Calicivirus VP1 Protein and Identification of Antigenic Epitopes

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