Purification of soybean agglutinin by affinity chromatography On sepharose‐N‐ϵ‐aminocaproyl‐β‐D‐galactopyranosylamine

On the presence of two types of subunit in soybean agglutinin

“…Soybean agglutinin was purified by affinity chromatography on Sepharose-N+-aminocaproyl-PD-galactopyranosylamine [7]. The 1ectin;was radioactively .…”

A simple method for the preparation of an affinity absorbent for soybean agglutinin using galactosamine and CH‐sepharose

“…The gel was then washed successively with 1 M NaCl, 1 M NaCl containcapacity for SBA (about 12 mg/ml gel) and like all ing 0.1 M Tris-HCl buffer (pH 8.6) 1 M NaCl consuch systems can be used repeatedly. The procedure taming 0.05 M-HCOONa buffer (pH 3.0) 1 M NaCl for the actual purification of SBA is essentially similar containing 0.1 M Tris-HCl buffer (pH 8.6), water and to that of Gordon et al [7,8] excepting that, with phosphate-buffered saline. The gel (6 ml) was then this batch of soybean meal, it was possible to omit the ready for use; if not immediately required it was storammonium sulphate fractionation and obtain a hunded in 2 M NaCl.…”

“…This is an analogue of N-acetylgalactosamine and is likely to have a greater affinity for SBA than is the derivative of galactose(caproylgalactosylamine) produced by Gordon et al [7,8]. The specificity of SBA is therefore directed towards N-acylgalactosamines in general rather than N-ucetylgalactosamine in particular.…”

“…SBA has been purified [3,4j and its properties have been investigated by Sharon et al [4,5] ; its agglutination reaction is specifically inhibited by N-acetyl-Dgaiactosamine and, to a lesser extent, by D-galactose [6]. Gordon et al [7,8] made an effective affinity system for the direct purification of SBA from an extract of soybean meal by linking N-e-aminocaproyl-flDgalactopyranosylamine to Sepharose. Although the purification system was straightforward and easy to use, the preparation of the affinity absorbent involves four steps, is fairly time-consuming and requires some of the facilities of an organic chemistry laboratory.…”

“…Although the purification system was straightforward and easy to use, the preparation of the affinity absorbent involves four steps, is fairly time-consuming and requires some of the facilities of an organic chemistry laboratory. We have found that an alternative affinity absorbent can easily be prepared by coupling galactosamine to CH-Sepharose (by a carbodiimide reaction in aqueous solution) with the production of a caproylamido linkage to C-2 of galactosamine, rather than a linkage to C-l of galactose in the system of Gordon et al [7,8]. The system that we have developed has a high affinity for SBA and the absorbed lectin can be eluted with Dgalactose.…”