Affinity fibre--a new support for rapid enzyme purification by high-performance liquid affinity chromatography.
A new type of support for chromatographic use has been developed. Nonporous quartz fibre, with a mean diameter of 0.5 micron, was silyated with mercaptopropyltrimethoxysilane. Tresyl chloride-activated dextran was covalently coupled to the SH groups on the fibre. Remaining active tresyl groups on the dextran were then coupled with an NAD derivative. The affinity fibre contained 0.3 mumol NAD derivative per g and was able to bind 15 mg of lactate dehydrogenase per g of fibre. The affinity fibre was used for large-scale purification of ox heart lactate dehydrogenase and its performance was compared with other commonly used chromatographic matrices. The operational capacity was found to be 1.0 g of pure lactate dehydrogenase per hour per 100 g opf fibre material (adsorption followed by salt elution). The affinity fibre was found to be particularly suitable for very rapid processing of large volumes of dilute enzyme solutions.
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004587:Electrophoresis, Agar Gel; D004798:Enzymes / Q000302:isolation & purification*; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification; D009243:NAD / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification; D011791:Quartz; D012822:Silicon Dioxide
D004798:Enzymes; D011506:Proteins; D009243:NAD; D011791:Quartz; D012822:Silicon Dioxide; D007770:L-Lactate Dehydrogenase