牛单核细胞趋化蛋白-1单克隆抗体的制备及应用

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中图分类号：S852.4 文献标志码：A 文章编号：1671-4652（2025）02-0001-08

DOI：10.16872/j.cnki.1671-4652.2025.02.001

ABSTRACT： In order to prepare monoclonal antibodyof bovine monocyte chemoattractant protein-1（BoMCP-1），the recombinant bacteria pGEX-6p-1-BoMCP-1and pET-30a-BoMCP-1 were constructedin this study.The purified prokaryotic expresson protein rHis-BoMCP-1 was used as an immunogen，and rGST-BoMCP-1 was used as a screening antigen. Among the mice immunized for three times，those with a positive serum titer of 1：150 000 or higher were choosed. MonOclonal antibodies were obtained throughcellfusionand monoclonal screning.Six strains of stably secreting specific BoMCP-1 monoclonal antibody wereobtained，named 3D10，4G9，4B6，1D8，3L4and 6F5.The affinityconstant reached （20 2.14×107 ， 5.12×107 ， 6.64×108 ， 8.42×107 ， 5.86×108 ， 3.61×107L⋅mol-1 . Flow cytometry showed that 4G9 had god reactivity with nativeBoMCP-1. In this study，bovine monocyte chemoatractant proteinl monoclonal antibody with strong specificity，highafinityand good natural reactivity was sucesfullyprepared.It laidafoundation forfurtherresearch on the pathogenesis and diagnostic application of MCP-1 in bovine epidemic diseases.

KEY WORDs： bovine； monocyte chemoattractant protein-l；monoclonal antibody； flow cytometry

趋化因子根据蛋白质N端保守的半胱氨酸残基的数量和间距分为CXC、CC、C和CX3C4个家族，其中大多数属于CC 和CXC亚家族。（剩余14859字）