KR910001043A - Polyethyleneimine Matrix for Affinity Chromatography - Google Patents

Abstract

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Description

Polyethyleneimine Matrix for Affinity Chromatography

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Claims (8)

(a) a support of the following general formula Silica-PrSi-PEI- (R) _X Wherein silica-PrSi-PEI (1) a) granular silica gel having an average particle diameter of about 1 to 200 microns and an average porosity of about 0 to 1000 Angstrom units, or b) an average particle diameter of about 1 to about 200 microns (2) Covalently bonded non-crosslinked polyethylene, the reaction product of granular controlled glass having an average porosity of 0 to about 100 Angstrom units with polyethylene amino propyl trimethoxy silane having an average molecular weight of about 400 to about 1800. A solid support of an imine-bonded phase, or (b) a weakly acidic carboxylation product containing from about 0.3 to about 1.2 carboxyl equivalents / gram of dibasic acid anhydride of a silica-PrSi-PEI solid support. Wherein, in the case of a non-acidic carboxylated support, R is any chemically reactive moiety capable of undergoing nucleophilic substitution at two distinct sites. As a group, R is covalently linked to the primary and secondary amino groups of the PEI at one such site while being useful and reactive for subsequent nucleophilic substitution under undenatured conditions by affinite chromatography ligands and thus stable under aqueous hydrolysis buffer conditions. Has other sites forming a covalent bond of 2, x is an integer such as less than or the total number of primary or secondary amino groups in the PEI moiety and in the case of a weakly acidic carboxylated support, R is the promoted nucleophilicity of the carboxyl hydroxyl As a residue of any chemically reactive moiety that can be substituted, it forms a covalent bond at the carboxyl carbon, thereby promoting a sufficiently nucleophilic site and is readily substituted at the carboxyl carbon by a nucleophilic functional group on the affinite chromatography ligand. Solid phase of less than or equal to the total number of carboxyl groups of the carboxylated PEI moiety Support. The solid phase support of claim 1, wherein the silica gel has an average particle diameter of about 3 to about 70 microns and an average porosity of about 50 to 100 Yongstrom units. The solid phase support according to claim 1 or 2, wherein R is selected from the group consisting of: An immobilized solid phase enzyme in which an enzyme is immobilized on an affiniter chromatography matrix solid phase support, wherein the solid phase support is a solid phase of any one of claims 1 to 3. (a) matrix Silica-PrSi-PEI (R) _X Wherein the silica-PrSi-PEI is (1) a) granular silica gel having an average particle diameter of about 1 to 200 microns and an average porosity of about 0 to 1000 Angstrom units, or b) about 1 to about 200 micron average particle diameter (2) Covalently bonded non-crosslinked polyethyleneimine bonds, the reaction product of granular controlled glass having an average porosity of 0 to 1000 Angstrom units with polyethylene amino propyl trimethoxy silane having an average molecular weight of about 400 to 1800. A solid support of a phase, or (b) a weakly acidic carboxylation product containing from about 0.3 to about 1.2 carboxyl milliequivalents per gram of dibasic acid anhydride of a silica-PrSi-PEI solid support. A bonded bond chromatographic affinity matrix consisting of a bond covalently bound to an affinity matrix, wherein in the case of a non-acidic carboxylated support R is a residue of any chemically reactive moiety that may be subjected to nucleophilic substitution at two separate sites, where R is covalently linked to the primary and secondary amino groups of PEI at one such site, while Has other sites that are useful for subsequent nucleophilic substitutions under denaturing conditions and are reactive to form a second covalent bond that is stable under aqueous hydrolysis buffer conditions and x is an integer such as less than or the total number of primary or secondary amino groups in the PEI moiety. And in the case of a weakly acidic carboxylated support, R is the residue of any chemically reactive moiety capable of facilitated nucleophilic substitution of the carboxyl hydroxyl, forming a covalent bond at the carboxyl carbon thereby promoting a sufficiently nucleophilic site and Easy substitution at the carboxyl carbon by nucleophilic functional groups on the nitichromatography ligand x is an integer recessed chromatography affinity matrix, such as a carboxyl group bonded gangeu less than the total number or the total number of parts of the carboxylated PEI. 6. The ligand-coupled chromatographic affinity matrix of claim 5, wherein the silica gel has an average particle diameter of about 3 to about 70 microns and an average workability of about 50 to 100 angstrom units. 7. The ligand bound chromatographic affinity matrix of claim 5 or 6, wherein R is selected from the group consisting of: A method of separating or purifying a substance from a solution by combining a substance in a solution with an affinity matrix having a ligand for a substance covalently bound to the affinity matrix, wherein the solid phase support of any one of claims 1 to 4 Separation or purification method comprising the use of a chemical conversion matrix. ※ Note: The disclosure is based on the initial application.