Lehrer R I, Rosenman M, Harwig S S, Jackson R, Eisenhauer P
Department of Medicine, UCLA Center for the Health Sciences 90024.
J Immunol Methods. 1991 Mar 21;137(2):167-73. doi: 10.1016/0022-1759(91)90021-7.
We developed two sensitive methods for identifying antimicrobial molecules in leukocytes and other tissues. One method uses a gel overlay technique and was designed to identify antimicrobial polypeptides in samples subjected to polyacrylamide gel electrophoresis. The other, a radial diffusion assay, allows multiple fractions obtained by chromatographic procedures to be tested for antimicrobial activity conveniently. When we used E. coli ML-35p or Salmonella typhimurium 14028S as test organisms in the radial diffusion assay, we routinely detected 5-10 ng of rabbit defensin NP-1 in 5 microliters of sample. With minor modifications, both methods can be applied to other organisms, including Gram-positive bacteria, several Candida species and Cryptococcus neoformans.
我们开发了两种灵敏的方法来鉴定白细胞和其他组织中的抗菌分子。一种方法采用凝胶覆盖技术，旨在鉴定经过聚丙烯酰胺凝胶电泳的样品中的抗菌多肽。另一种方法是径向扩散测定法，可方便地检测通过色谱程序获得的多个组分的抗菌活性。当我们在径向扩散测定法中使用大肠杆菌ML-35p或鼠伤寒沙门氏菌14028S作为测试生物时，我们通常能在5微升样品中检测到5-10纳克的兔防御素NP-1。稍作修改后，这两种方法均可应用于其他生物，包括革兰氏阳性菌、几种念珠菌属物种和新型隐球菌。
