Fernández L A, Sola I, Enjuanes L, de Lorenzo V
Centro Nacional de Biotecnología del Consejo Superior de Investigaciones Científicas (CSIC), Campus de Cantoblanco, 28049 Madrid, Spain.
Appl Environ Microbiol. 2000 Nov;66(11):5024-9. doi: 10.1128/AEM.66.11.5024-5029.2000.
A simple method for the nontoxic, specific, and efficient secretion of active single-chain Fv antibodies (scFvs) into the supernatants of Escherichia coli cultures is reported. The method is based on the well-characterized hemolysin transport system (Hly) of E. coli that specifically secretes the target protein from the bacterial cytoplasm into the extracellular medium without a periplasmic intermediate. The culture media that accumulate these Hly-secreted scFv's can be used in a variety of immunoassays without purification. In addition, these culture supernatants are stable over long periods of time and can be handled basically as immune sera.
报道了一种将活性单链Fv抗体(scFv)无毒、特异性且高效地分泌到大肠杆菌培养上清液中的简单方法。该方法基于大肠杆菌特征明确的溶血素转运系统(Hly),该系统可将靶蛋白从细菌细胞质特异性分泌到细胞外培养基中,无需周质中间体。积累这些经Hly分泌的scFv的培养基无需纯化即可用于多种免疫测定。此外,这些培养上清液在很长一段时间内都很稳定,基本上可以像免疫血清一样处理。