Polysaccharide matrices for use as adsorbents in affinity chromatography techniques
OwnerPedro Cuatrecasas, Indu Parikhinventors
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Abstract
Polysaccharide matrices, e.g., agarose, for use in certain affinity chromatography procedures, are activated with sodium metaperiodate, derivatized with a symmetrical dihydrazide and thence reductively stabilized, preferably with sodium cyanoborohydride. Moreover, intermediate Schiff bases formed from the NaIO.sub.4 -oxidized polysaccharide are selectively, reductively stabilized with said sodium cyanoborohydride.
Claims19
In the method for the activation of a polysaccharide matrix for use as an adsorbent in affinity chromatography techniques, comprising activating the said polysaccharide by oxidizing the cis-vicinal hydroxyl groups thereof with sodium metaperiodate to generate aldehyde functions, converting said aldehydic functions into Schiff bases and thence reductively stabilizing the said resultant Schiff bases, the improvement which comprises the reductive stabilization of said intermediate Schiff bases with sodium cyanoborohydride at a pH between 6 and 6.5.
The method for the preparation of a hydrazide-polysaccharide derivative for use as an adsorbent in affinity chromatography techniques, which comprises activating a polysaccharide matrix by oxidizing the cis-vicinal hydroxyl groups thereof with sodium metaperiodate to generate aldehyde functions, reacting said periodate-oxidized polysaccharide with a bifunctional, symmetrical dihydrizide to convert the same to the corresponding hydrazido/hydrazone derivative, and thence reductively stabilizing said hydrazido/hydrazone derivative with sodium cyanoborohydride at a pH between 6 and 6.5 to produce an unsemetric hydrazide.
Dependencies are inferred from the claim text.