McQuade T J, Pitts T W, Tarpley W G
Cancer and Infectious Diseases Research, Upjohn Company, Kalamazoo, MI 49001.
Biochem Biophys Res Commun. 1989 Aug 30;163(1):172-6. doi: 10.1016/0006-291x(89)92116-5.
We developed a particle concentration fluorescent immunoassay to quantify the binding in solution of the human immunodeficiency virus (HIV) external glycoprotein (gp120) to soluble CD4 (sCD4). The assay is rapid (1 hr), quantitative, and requires as little as 0.1 pmole of gp120 per evaluation. We find that gp120, purified from recombinant baculovirus infected insect cells, is suitable for the assay. Moreover, sCD4s obtained either from recombinant E. coli or mammalian cells, consisting of the N-terminal two domains (about 180 amino acids) as well as linked to the active regions of Pseudomonas exotoxin A, bind gp120 similarly.
我们开发了一种颗粒浓度荧光免疫测定法，用于定量人免疫缺陷病毒（HIV）外膜糖蛋白（gp120）与可溶性CD4（sCD4）在溶液中的结合。该测定法快速（1小时）、定量，每次评估仅需0.1皮摩尔的gp120。我们发现，从重组杆状病毒感染的昆虫细胞中纯化的gp120适用于该测定法。此外，从重组大肠杆菌或哺乳动物细胞中获得的sCD4，由N端的两个结构域（约180个氨基酸）组成，并与外毒素A的活性区域相连，它们与gp120的结合情况相似。
