Sagisaka K, Hamada C
Tohoku J Exp Med. 1975 Jul;116(3):297-8. doi: 10.1620/tjem.116.297.
Sialoprotein possessing blood group activity has been usually extracted with organic solvents from red cell membrane. To obtain the blood group substance under milder condition, the author presented carboxymethyl (CM) cellulose column chromatography with eluents of phosphate buffer containing 1% Brij 35. Red cell stroma was applied onto the column with an equal volume of 0.0175 M phosphate buffer pH 6.3 and one-fifth volume of 0.05% 2-mercaptoethanol in 1% Brij 35. Separation of the substance was performed by stepwise elution with three kinds of phosphate buffers. Serological assay showed that only the first fraction eluted with 1% Brij 35 in 0.0175 M phosphate buffer pH 6.3 and ABH and MN blood group activities and the other fractions obtained with buffers of higher molarities had not such activities. Sialic acid was also detected only in the first fraction, and the majority of proteins of the stroma was found in the other fractions.
具有血型活性的唾液酸蛋白通常是用有机溶剂从红细胞膜中提取的。为了在更温和的条件下获得血型物质，作者提出了用含1% 聚氧乙烯月桂醚35的磷酸盐缓冲液作为洗脱剂的羧甲基（CM）纤维素柱色谱法。将红细胞基质用等体积的pH 6.3的0.0175 M磷酸盐缓冲液和五分之一体积的含1% 聚氧乙烯月桂醚35的0.05% 2-巯基乙醇上样到柱上。用三种磷酸盐缓冲液分步洗脱进行物质分离。血清学检测表明，只有在pH 6.3的0.0175 M磷酸盐缓冲液中用1% 聚氧乙烯月桂醚35洗脱的第一部分具有ABH和MN血型活性，而用更高摩尔浓度缓冲液获得的其他部分没有这种活性。唾液酸也仅在第一部分中检测到，并且基质中的大多数蛋白质存在于其他部分中。
