Marcet Sánchez R, Méndez Pérez I, Castaño J C, Otero Alfaro O, Lastre González M, Pérez Martín O, Sarracent Pérez J
Universidad del Quindío, Colombia.
Rev Cubana Med Trop. 1999 Jan-Apr;51(1):33-7.
In order to determine the IgA levels in secretions, the authors engaged themselves to obtain monoclonal antibodies against this immunoglobulin and to show their possible utilization in the development of quantitation methods. 6 monoclonal antibodies were obtained: one against the secretory component (SC) and five against the heavy-chain (HC) of IgA. IgA purified from human colostrum was used as immunogen (HC). The immunoglobulin isotypes (IgG2a and IgG1) were determined in 2 monoclonal antibodies, one against SC and the other against HC. Polyacrylamide gel electrophoresis was performed in the presence of sodium dodecyl sulphate and b-mercapto-ethanol. Immunotransference was also carried out to determine their specificity. Secretory IgA was detected in saliva specimens. The anti-SC monoclonal antibody obtained in-the laboratory of the "Pedro Kourí" Institute of Tropical Medicine (IPK) was compared with a commercial one (Sigma Chemical Co.), and a similar behaviour was observed. The monoclonal antibody against SC (IPK) was used to detect IgA specific to LPS ogawa in persons vaccinated with an attenuated strain of Vibrio cholerae. Marked differences were found between the levels before and after immunization. The anti-HC monoclonal antibody was utilized to make an ELISA-like simulated system. The detection limit was 9.89 ng/mL.
为了测定分泌物中的IgA水平，作者致力于获取针对这种免疫球蛋白的单克隆抗体，并展示其在定量方法开发中的可能应用。获得了6种单克隆抗体：一种针对分泌成分（SC），五种针对IgA的重链（HC）。从人初乳中纯化的IgA用作免疫原（HC）。在两种单克隆抗体中测定了免疫球蛋白同种型（IgG2a和IgG1），一种针对SC，另一种针对HC。在十二烷基硫酸钠和β-巯基乙醇存在下进行聚丙烯酰胺凝胶电泳。还进行了免疫转移以确定其特异性。在唾液标本中检测到分泌型IgA。将在“佩德罗·库里”热带医学研究所（IPK）实验室获得的抗SC单克隆抗体与市售抗体（西格玛化学公司）进行比较，观察到类似的行为。使用抗SC单克隆抗体（IPK）检测接种霍乱弧菌减毒株的人群中针对小川型脂多糖的特异性IgA。发现免疫前后的水平存在明显差异。利用抗HC单克隆抗体制备了类似ELISA的模拟系统。检测限为9.89 ng/mL。
