Heimerman Mallory E, Murgia Maria V, Wu Ping, Lowe Andre D, Jia Wei, Rowland Raymond R
Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS (Heimerman, Murgia, Rowland).
Foreign Animal Disease Diagnostic Laboratory, National Veterinary Services Laboratories, Animal and Plant Health Inspection Services, U.S. Department of Agriculture, Plum Island Animal Disease Center, New York, NY (Wu, Lowe, Jia).
J Vet Diagn Invest. 2018 May;30(3):406-412. doi: 10.1177/1040638717753966. Epub 2018 Jan 12.
Protein p72 is the major capsid protein of African swine fever virus (ASFV) and is an important target for test and vaccine development. Monoclonal antibodies (mAbs) were prepared against a recombinant antigenic fragment, from amino acid (aa) 20-303, expressed in baculovirus. A total of 29 mAbs were recovered and tested by immunofluorescent antibody (IFA) staining on ASFV Lisbon-infected Vero cells. Six antibodies were IFA-positive and selected for further characterization. Epitope mapping was performed against overlapping polypeptides expressed in E. coli and oligopeptides. Based on oligopeptide recognition, the mAbs were divided into 4 groups: mAb 85 (aa 165-171); mAbs 65-3 and 6H9-1 (aa 265-280); mAbs 8F7-3 and 23 (aa 280-294); and mAb 4A4 (aa 290-303). All mAbs were located within a highly conserved region in p72. This panel of antibodies provides the opportunity to develop new assays for the detection of ASFV antibody and antigen.
p72蛋白是非洲猪瘟病毒（ASFV）的主要衣壳蛋白，也是检测和疫苗研发的重要靶点。针对杆状病毒中表达的氨基酸（aa）20 - 303的重组抗原片段制备了单克隆抗体（mAb）。共获得29种单克隆抗体，并通过免疫荧光抗体（IFA）染色在感染了ASFV里斯本株的Vero细胞上进行检测。六种抗体IFA呈阳性，并被选作进一步鉴定。针对在大肠杆菌中表达的重叠多肽和寡肽进行表位作图。基于寡肽识别，单克隆抗体被分为4组：单克隆抗体85（aa 165 - 171）；单克隆抗体65 - 3和6H9 - 1（aa 265 - 280）；单克隆抗体8F7 - 3和23（aa 280 - 294）；以及单克隆抗体4A4（aa 290 - 303）。所有单克隆抗体都位于p72的一个高度保守区域内。这组抗体为开发检测ASFV抗体和抗原的新检测方法提供了机会。
