利用链霉亲和素-生物素相互作用制备用于蛋白质补体依赖性脂质体免疫测定的试剂：检测Latrotoxin毒素。
The use of streptavidin-biotin interaction for preparation of reagents for complement-dependent liposome immunoassay of proteins: detection of latrotoxin.
作者信息
Pashkov V N, Tsurupa G P, Griko N B, Skopinskaya S N, Yarkov S P
机构信息
Branch of the M. M. Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow Region, Russia.
出版信息
Anal Biochem. 1992 Dec;207(2):341-7. doi: 10.1016/0003-2697(92)90022-y.
We have developed liposome sensitization by a protein, latrotoxin (LT), using immobilization of biotinylated LT via streptavidin with biotinylated phosphatidylethanolamine contained in liposomes. The use of such liposomes in the complement-dependent homogeneous liposome immune lysis assay (LILA) has allowed us to detect in the test sample as little as 2 micrograms/ml of polyclonal and 50-100 ng/ml of monoclonal IgG and IgM antibodies to LT. LT concentration in solution was determined by inhibition of immune lysis by free LT. The sensitivity of the LT assay varied from 1 x 10(-9) to 5-50 x 10(-9) M when antiserum (polyclonal antibodies) and monoclonal antibodies to LT were correspondingly used. The results show that a streptavidin-biotin spacer can be used to immobilize protein antigens on liposomes for a subsequent application in LILA. The suggested technique greatly simplifies the sensitization procedure and extends the applicability of the LILA.
我们通过一种蛋白质——黑寡妇毒素（LT）开发了脂质体致敏方法，即利用链霉亲和素将生物素化的LT与脂质体中含有的生物素化磷脂酰乙醇胺固定在一起。在补体依赖的均相脂质体免疫裂解试验（LILA）中使用这种脂质体，使我们能够在测试样品中检测到低至2微克/毫升的多克隆抗体以及50 - 100纳克/毫升的针对LT的单克隆IgG和IgM抗体。溶液中的LT浓度通过游离LT对免疫裂解的抑制作用来确定。当相应地使用抗血清（多克隆抗体）和针对LT的单克隆抗体时，LT检测的灵敏度在1×10⁻⁹至5 - 50×10⁻⁹ M之间变化。结果表明，链霉亲和素 - 生物素间隔物可用于将蛋白质抗原固定在脂质体上，以便随后应用于LILA。所建议的技术极大地简化了致敏程序并扩展了LILA的适用性。
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