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以开发酶联免疫吸附测定试剂盒为最终目标的针对牛传染性胸膜肺炎抗原的单克隆抗体的研发与评估

Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit.

作者信息

Ramathudi-Dunbar Lorato, Awosanya Emmanuel, Charles Sanne Bodjo, Chitsungo Ethel, Moustapha Boukary Cisse Rahamatou, Nwankpa Nick, Gelaw Hassen, Tessema Yebechaye, Melesse A Gelagay, Rayson Sanga Richard, Bright Adorbley, Baziki Jean de Dieu

机构信息

Pan African University Life Sciences Institute Including Health and Agriculture (PAULESI), University of Ibadan, Ibadan, Nigeria.

African Union Pan African Veterinary Vaccine Centre (AU-PANVAC), P.O. Box 1746, Debre Zeit, Ethiopia.

出版信息

Vet Med Int. 2024 May 7;2024:6901355. doi: 10.1155/2024/6901355. eCollection 2024.

Abstract

Contagious bovine pleuropneumonia (CBPP) is an infectious and contagious bacterial respiratory disease that affects cattle with significant economic losses to the African animal industry. The use of ELISA kits based on monoclonal antibodies (mAbs) will aid in quick and precise diagnosis of CBPP, contributing to disease control and prevention in cattle. Thus, this research aims to develop and evaluate monoclonal antibodies against CBPP (T1/44) antigen for use in ELISA kits for CBPP diagnosis. Hybridoma technology was used to develop monoclonal antibodies that recognize and bind to the CBPP (T1/44) antigen. The antibody-secreting hybridomas were produced after immunizing mice with purified CBPP antigens. The hybridomas were screened for high sensitivity, specificity, and liking to the antigen. The selected mAbs were assessed for sensitivity and specificity against CBPP antigen using different immunoassays, dot-blot, ELISA, and mouse mAb isotyping. The monoclonal antibodies were profoundly specific, with a higher hindrance to CBPP antigen (<0.50 OD) while lacking cross-reactivity to other antigens. The monoclonal antibodies could distinguish CBPP antigen at low concentrations, showing their high sensitivity (>80% PI). The isotyped mAbs of intrigued appeared to have a place in the IgG class. These identified monoclonal antibodies can be utilized to develop an ELISA kit for CBPP diagnosis, which would give a fast, precise, and cost-effective strategy for screening and checking CBPP in cattle herds.

摘要

牛传染性胸膜肺炎(CBPP)是一种传染性细菌性呼吸道疾病,会感染牛群,给非洲畜牧业造成重大经济损失。基于单克隆抗体(mAb)的ELISA试剂盒的使用将有助于快速、准确地诊断CBPP,有助于牛群疾病的控制和预防。因此,本研究旨在开发和评估针对CBPP(T1/44)抗原的单克隆抗体,用于CBPP诊断的ELISA试剂盒。采用杂交瘤技术开发能识别并结合CBPP(T1/44)抗原的单克隆抗体。在用纯化的CBPP抗原免疫小鼠后产生分泌抗体的杂交瘤。对杂交瘤进行高灵敏度、特异性和与抗原亲和力的筛选。使用不同的免疫测定法(斑点印迹法、ELISA和小鼠单克隆抗体分型)评估所选单克隆抗体对CBPP抗原的敏感性和特异性。这些单克隆抗体具有高度特异性,对CBPP抗原的阻滞作用较高(<0.50 OD),同时与其他抗原无交叉反应。单克隆抗体能够在低浓度下区分CBPP抗原,显示出它们的高灵敏度(>80% PI)。感兴趣的分型单克隆抗体似乎属于IgG类。这些鉴定出的单克隆抗体可用于开发用于CBPP诊断的ELISA试剂盒,这将为牛群中CBPP的筛查和检测提供一种快速、准确且经济高效的策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cb54/11093687/497180fdb890/VMI2024-6901355.001.jpg

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