Elution of tightly bound solutes from concanavalin A Sepharose. Factors affecting the desorption of cottonmouth venom glycoproteins.
Some glycoproteins bind so tightly to concanavalin A Sepharose that common desorption techniques are ineffective, so a systematic exploration of factors affecting desorption of cottonmouth venom glycoproteins was undertaken. Glycoprotein desorption is greatly improved by introducing up to four pauses of 5-10 min duration into the elution step. Eluent concentrations above 250 mM methylglucoside or 500 mM methyl-mannoside reduced glycoprotein desorption. Eluent NaCl diminished glycoprotein desorption. Most venom glycoproteins desorb more readily as pH diminishes from 6.0 to 4.0, but phosphodiesterase shows the opposite pattern. Eluents recommended by the supplier for desorbing solutes or for column cleaning were ineffectual.
Soper
Anastasiya S
AS
Department of Chemistry, Norfolk State University, 700 Park Avenue, Norfolk, VA 23504, USA.
Aird
Steven D
SD
D000327:Adsorption; D017836:Agkistrodon; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003435:Crotalid Venoms / Q000302:isolation & purification*; D006023:Glycoproteins / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D010727:Phosphoric Diester Hydrolases / Q000302:isolation & purification; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry
C031442:Agkistrodon venoms; D003435:Crotalid Venoms; D006023:Glycoproteins; C028326:concanavalin A-sepharose; D012685:Sepharose; D010727:Phosphoric Diester Hydrolases