Pyle S W, Bess J W, Robey W G, Fischinger P J, Gilden R V, Arthur L O
Program Resources, Inc., Frederick Cancer Research Facility (NCI-FCRF), MD 21701.
AIDS Res Hum Retroviruses. 1987;3(4):387-400. doi: 10.1089/aid.1987.3.387.
The outer envelope glycoprotein, gp120, has been purified from large volumes (greater than 100 liters) of HTLV-IIIB-infected H9 cell culture fluids using immunoaffinity chromatography resins prepared from immunoglobulins of AIDS patients plasma. By using a single-step immunoaffinity purification, between 7 and 28 micrograms of gp120 was recovered from each liter of culture fluid which represented between 60 and 95% of the total envelope glycoprotein present in the fluid. Envelope glycoprotein in culture media was concentrated more than 10,000 times over the starting material. The water-soluble gp120, containing trace contaminating proteins, was purified to apparent homogeneity by preparative polyacrylamide gel electrophoresis (PAGE). Envelope glycoprotein purified from culture fluids was immunogenic in laboratory animals in both native and PAGE-purified forms and was reactive with AIDS patient sera in immunoassays.
外膜糖蛋白gp120是利用从艾滋病患者血浆免疫球蛋白制备的免疫亲和层析树脂，从大量（超过100升）感染HTLV-IIIB的H9细胞培养液中纯化得到的。通过单步免疫亲和纯化，每升培养液可回收7至28微克的gp120，占培养液中总包膜糖蛋白的60%至95%。培养基中的包膜糖蛋白比起始原料浓缩了10000倍以上。含有微量污染蛋白的水溶性gp120通过制备性聚丙烯酰胺凝胶电泳（PAGE）纯化至表观均一。从培养液中纯化的包膜糖蛋白以天然形式和PAGE纯化形式在实验动物中具有免疫原性，并且在免疫测定中与艾滋病患者血清发生反应。
