Rantamäki L K, Müller H P
Department of Microbiology and Epizootology, College of Veterinary Medicine, Helsinki, Finland.
Vet Immunol Immunopathol. 1995 Mar;45(1-2):115-26. doi: 10.1016/0165-2427(94)05329-q.
The application of stabilised streptococcal cells for the purification of both immunoglobulin 1 (IgG1) and IgG2 subclasses from goat sera was evaluated. Guanidinium chloride extracted, lyophilised cells of the Lancefield Group C Streptococcus dysgalactiae Sc1 strain showed strong binding to goat IgG, reaching a capacity of approximately 1.4 mg IgG per 100 mg cells (dry weight). The IgG preparation obtained was of high quality. In immunoelectrophoretic analysis the preparation appeared to consist of pure IgG, whereas the high pressure liquid chromatography (HPLC) gel filtration and immunoblot analyses showed a very slight contamination (less than 1.3% of the total probe) with alpha 2-macroglobulin. The presence of both IgG subclasses in the preparation was verified by HPLC ion exchange chromatography. The adsorption procedure proved to be efficient and easy to perform without advanced technical equipment and the cells were reusable. As these streptococcal cells bind both IgG subclasses, this method presents an economical way for the small scale purification of goat IgG. Additionally the streptococcal cells may conveniently substitute Protein A bearing Staphylococcus aureus cells in various immunological assays.
评估了稳定化链球菌细胞用于从山羊血清中纯化免疫球蛋白1(IgG1)和IgG2亚类的应用。用氯化胍提取、冻干的C群马链球菌(Streptococcus dysgalactiae)Sc1菌株细胞对山羊IgG显示出强结合力,每100 mg细胞(干重)结合IgG的能力约为1.4 mg。所获得的IgG制剂质量很高。在免疫电泳分析中,该制剂似乎由纯IgG组成,而高压液相色谱(HPLC)凝胶过滤和免疫印迹分析显示存在极少量(占总探针的不到1.3%)的α2-巨球蛋白污染。通过HPLC离子交换色谱法验证了制剂中两种IgG亚类的存在。吸附过程证明是高效的,且无需先进技术设备即可轻松进行,细胞可重复使用。由于这些链球菌细胞能结合两种IgG亚类,该方法为小规模纯化山羊IgG提供了一种经济的方式。此外,在各种免疫测定中,链球菌细胞可以方便地替代携带蛋白A的金黄色葡萄球菌细胞。