| Abstract: | Abstract Concanavalin A- Sepharose affinity chromatography is a powerful tool for isolation or purification of peripheral or integral membrane proteins or other glycoproteins. The insecticidal crystal toxin from Bacillus thuringiensis subsp. kurstaki is a glycoprotein containing “high-mannose” or “hybrid”-type sugar chains. The protein has a high binding affinity for concanavalin A lectin and could not be eluted even with 0.5M methyl α-D-mannopyranoside. Nonspecific elution with 0.03% SDS coeluted the matrix con A with bound protein. Experimental results indicated that con A leaching is mainly because of inclusion of detergents in buffer systems and may not be directly related to the nature of the sample protein. 2 Abbreviations used: Con A: concanavalin A, SDS: sodium dodecyl sulfate, SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis, MeG: methyl α-D-glucopyranoside, MeM: methyl α-D-mannopyranoside |
