Yamamoto S, Tagata K, Ishikawa Y, Fujise H, Nagahata H, Yamada M, Sakano T, Morimatsu M, Naiki M
Laboratory of Immunology, Faculty of Environmental Health, Azabu University, Kanagawa, Japan.
Vet Res Commun. 1992;16(4):265-72. doi: 10.1007/BF01839325.
A method is described for preparing latex particles sensitized with IgG antibody (IgG-sensitized latex) applicable to the slide reversed passive agglutination (RPLA) test. Soap-free latex (latex) was sensitized with IgG which had been isolated from rabbit anti-bovine lactoferrin serum using protein A Sepharose CL 4B. Unadsorbed protein-binding sites on the surface of latex were blocked with bovine serum albumin (BSA). IgG-sensitized latex that gave better agglutination in RPLA could be selectively obtained by centrifugation at 19 900g for 15 min in 0.01 mol/L glycine buffer (pH 7.3; specific gravity 1.042) containing 3% NaCl, 5% saccharose and 2% choline chloride. By dispersing this IgG-sensitized latex in 0.01 mol/L glycine buffer (pH 7.3) containing 1-2% BSA, a uniformly suspended, highly reactive, readily agglutinable preparation was obtained.
描述了一种制备适用于玻片反向被动凝集（RPLA）试验的用IgG抗体致敏的乳胶颗粒（IgG致敏乳胶）的方法。使用蛋白A琼脂糖凝胶CL 4B从兔抗牛乳铁蛋白血清中分离出的IgG致敏无皂乳胶（乳胶）。乳胶表面未吸附的蛋白质结合位点用牛血清白蛋白（BSA）封闭。通过在含有3%氯化钠、5%蔗糖和2%氯化胆碱的0.01mol/L甘氨酸缓冲液（pH 7.3；比重1.042）中以19900g离心15分钟，可以选择性地获得在RPLA中凝集效果更好的IgG致敏乳胶。通过将这种IgG致敏乳胶分散在含有1-2% BSA的0.01mol/L甘氨酸缓冲液（pH 7.3）中，得到了均匀悬浮、高反应性、易于凝集的制剂。
