Nollens Hendrik H, Green Linda G, Duke Diane, Walsh Michael T, Chittick Beth, Gearhart Scott, Klein Paul A, Jacobson Elliott R
Marine Mammal Health Program, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
J Vet Diagn Invest. 2007 Sep;19(5):465-70. doi: 10.1177/104063870701900502.
Antibodies directed against species-specific immunoglobulin G (IgG) have a broad range of applications in serologic and immunologic research and in the development of clinical assays. Validated anti-IgG antibodies for marine mammal species are in short supply. The objective of this study was to produce and validate antibodies with specificity for IgG of the common bottlenose dolphin (Tursiops truncatus). Bottlenose dolphin IgG was purified using protein G. Two mouse monoclonal antibodies and a rabbit polyclonal antibody were developed from mice and rabbits immunized with bottlenose dolphin IgG. The specificity of the monoclonal antibodies and the polyclonal antibody for bottlenose dolphin IgG was first verified by Western blot analysis and enzyme-linked immunosorbent assay (ELISA). For further validation, both monoclonal antibodies and the polyclonal antibody were incorporated in an indirect ELISA for the detection of the immune response of bottlenose dolphins to a vaccine antigen. Three bottlenose dolphins were immunized with a commercial Erysipelothrix rhusiopathiae vaccine, and serial blood samples were collected from all dolphins for measurement of levels of circulating antibodies. Seroconversion was observed in all 3 dolphins by use of both monoclonal antibodies and the polyclonal antibody. Circulating antibodies were detectable as early as 6 days after immunization in 1 dolphin. Peak antibody levels were detected 14 days after the immunization. The ability to detect seroconversion in all 3 immunized bottlenose dolphins firmly establishes the specificity of the monoclonal antibodies and the polyclonal antibody for IgG of the common bottlenose dolphin.
针对物种特异性免疫球蛋白G（IgG）的抗体在血清学和免疫学研究以及临床检测方法的开发中具有广泛应用。经过验证的针对海洋哺乳动物物种的抗IgG抗体供应短缺。本研究的目的是制备并验证对宽吻海豚（瓶鼻海豚，Tursiops truncatus）IgG具有特异性的抗体。利用蛋白G纯化宽吻海豚IgG。用宽吻海豚IgG免疫小鼠和兔子，分别制备了两种小鼠单克隆抗体和一种兔多克隆抗体。首先通过蛋白质印迹分析和酶联免疫吸附测定（ELISA）验证单克隆抗体和多克隆抗体对宽吻海豚IgG的特异性。为了进一步验证，将单克隆抗体和多克隆抗体均用于间接ELISA，以检测宽吻海豚对疫苗抗原的免疫反应。用市售的猪丹毒杆菌疫苗免疫三只宽吻海豚，并从所有海豚采集系列血样以测量循环抗体水平。使用单克隆抗体和多克隆抗体均在所有三只海豚中观察到血清转化。在一只海豚中，免疫后最早6天就能检测到循环抗体。免疫后14天检测到抗体水平峰值。在所有三只免疫的宽吻海豚中均能检测到血清转化，这有力地证实了单克隆抗体和多克隆抗体对宽吻海豚IgG的特异性。
