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使用抗纤溶酶原激活酶LV-PA的抗体对矛头蝮蛇毒进行特异性鉴定。

Specific identification of Lachesis muta muta snake venom using antibodies against the plasminogen activator enzyme, LV-PA.

作者信息

Felicori Liza F, Chávez-Olórtegui Carlos, Sánchez Eladio F

机构信息

Centro de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, 30510-010 Belo Horizonte, MG, Brazil.

出版信息

Toxicon. 2005 May;45(6):803-6. doi: 10.1016/j.toxicon.2004.12.003.

Abstract

Sandwich-type enzyme linked immunosorbent assays (ELISA) were developed to detect Lachesis muta muta (bushmaster) snake venom using antibodies against the plasminogen activator enzyme (LV-PA). Antibodies to LV-PA were obtained by immunization of one rabbit with the purified enzyme. The IgG fraction was purified from rabbit blood in a single step on a column of Sepharose-L. m. muta venom and used to coat the microtiter plates. The specificity of the assay was demonstrated by its capacity to correctly discriminate between the circulating antigens in mice that were experimentally inoculated with L. m. muta venom from those in mice inoculated with venoms from Bothrops atrox, B. brazili, B. castelnaudi, Bothriopsis taeniata, B. bilineata, Crotalus durissus ruruima and the antigenic Bothrops (AgB) and Crotalus (AgC) pools venoms used to produce Bothropic and Crotalic antivenoms at Fundacao Ezequiel Dias (FUNED). Measurable absorbance signals were obtained with 1.5 ng of venom per assay. The ELISA was used to follow the kinetic distribution of antigens in experimentally envenomed mice.

摘要

开发了夹心型酶联免疫吸附测定法(ELISA),以使用抗纤溶酶原激活酶(LV-PA)的抗体来检测矛头蝮蛇毒。通过用纯化的酶免疫一只兔子获得针对LV-PA的抗体。IgG组分在Sepharose-L.m.muta毒液柱上一步从兔血中纯化出来,并用于包被微量滴定板。该测定法的特异性通过其正确区分实验接种了L.m.muta毒液的小鼠体内的循环抗原与接种了巴西矛头蝮、巴西矛头蝮、卡斯泰尔诺迪矛头蝮、带纹竹叶青、双线竹叶青、杜氏响尾蛇uruima以及用于在埃泽奎埃尔·迪亚斯基金会(FUNED)生产抗巴西矛头蝮和抗响尾蛇抗蛇毒血清的抗原性巴西矛头蝮(AgB)和响尾蛇(AgC)混合毒液的小鼠体内的循环抗原的能力得以证明。每次测定使用1.5 ng毒液可获得可测量的吸光度信号。ELISA用于追踪实验性中毒小鼠体内抗原的动力学分布。

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