Singh-Bist A, Maheux P, Azhar S, Chen Y D, Komaromy M C, Kraemer F B
Department of Medicine, Stanford University School of Medicine, CA 94305, USA.
Life Sci. 1995;57(18):1709-15. doi: 10.1016/0024-3205(95)02150-h.
Antibodies generated against specific proteins are useful tools for studying the physiology and cell biology of the protein of interest. Although antibodies have been successfully generated against lipoprotein lipase (LPL) and used to elucidate many aspects of its biology, there have been problems with the specificity, affinity and availability of these antibodies. To circumvent these problems, we have expressed a portion of human LPL as a bacterial fusion protein. The human LPL bacterial fusion protein was utilized to generate polyclonal antibodies in rabbits that recognize intact human, rat and bovine LPL. Using these antibodies, it was possible to demonstrate a direct correlation between LPL mass and LPL activity from different samples of human post-heparin plasma. In addition, these antibodies were used to develop an ELISA for the measurement of LPL in tissue or plasma. This is a useful means for obtaining polyclonal antibodies to LPL in sufficient quantity and without contaminating mammalian proteins.
针对特定蛋白质产生的抗体是研究目标蛋白质生理学和细胞生物学的有用工具。尽管已经成功产生了针对脂蛋白脂肪酶（LPL）的抗体，并用于阐明其生物学的许多方面，但这些抗体在特异性、亲和力和可获得性方面存在问题。为了规避这些问题，我们将人LPL的一部分表达为细菌融合蛋白。利用人LPL细菌融合蛋白在兔中产生多克隆抗体，这些抗体可识别完整的人、大鼠和牛LPL。使用这些抗体，有可能证明来自不同人肝素后血浆样本的LPL质量与LPL活性之间存在直接相关性。此外，这些抗体被用于开发一种用于测量组织或血浆中LPL的ELISA。这是一种获得足够数量的针对LPL的多克隆抗体且不污染哺乳动物蛋白质的有用方法。
