Polyethyleneimine matrixes for affinity chromatography

• US 5,092,992 A
• Filed: 05/17/1991
• Issued: 03/03/1992
• Est. Priority Date: 06/07/1989
• Status: Expired due to Term

• Current Assignee: Avantor Performance Materials, Inc. (Avantor, Inc.)
• Original Assignee: J.T. Baker, Inc. (Avantor, Inc.)
• Inventors: Crane, Laura J., Kakodkar, Sunil V.
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US07/703,039
• Time in Patent Office: 291 Days
• Field of Search: 210/635, 210/656, 210/198.2, 210/502.1, 502/401, 502/402, 530/413, 530/417
• US Class Current: 210/198.2
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/3257 the functional group or the linking, spacer or anchoring group as a whole comprising at least one of the heteroatoms nitrogen, oxygen or sulfur together with at least one silicon atom, these atoms not being part of the carrier as such
• CPC Class Codes: B01J 20/3263 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. an heterocyclic or heteroaromatic structure
• CPC Class Codes: B01J 20/328 Polymers on the carrier being further modified

Abstract

Affinity chromatography matrixes and immobilized enzymes are provided in which the solid support is a covalently bound, non-crosslinked polyethyleneimine bonded phase.

Claims

1. A solid phase support suitable for use in a column selected from the group consisting of:

(a) a support of the general formula

space="preserve" listing-type="equation">Silica--PrSi--PEI--R).sub.x

wherein Silica--PrSi--PEI is a covalently bound, non-crosslinked polyethyleneimine bonded phase solid support which is the reaction product of (1) a) particulate silica gel having an average particle diameter of from about 1 to 200 microns and an average pore size of from about 0 to 1,000 Angstrom units, or b) particulate controlled pore glass having an average particle diameter of from about 1 to about 200 microns and an average pore size of from about 0 to about 1,000 Angstrom units, with (2) polyethyleneiminopropyl trimethoxy silane having an average molecular weight of from about 400 to about 1,800, or

(b) the weakly acidic carboxylated product of the Silica--PrSi--PEI--r)_x solid support with a dibasic acid anhydride, said carboxylated product containing from about 0.3 to about 1.2 carboxyl milliequivalents per gram, and wherein R in the case of the non-weakly acidic carboxylated support is the residue of any chemically reactive moiety capable of undergoing nucleophilic substitution at two separate sites, such that R becomes covalently linked to both primary and secondary amino groups of the PEI at one such site while having the other site available and reactive for subsequent nucleophilic substitution under non-denaturing conditions by an affinity chromatography ligand to form a second covalent bond stable under aqueous hydrolytic buffer conditions and x is a positive integer less than or equal to the total number of primary or secondary amino groups in the PEI moiety, and in the case of the weakly acidic caboxylated support R is the residue of any chemically reactive moiety capable of facilitated nucleophilic displacement of the carboxyl hydroxyl to form a covalent bond at the carboxyl carbon, creating thereby a sufficiently electrophilic site, so as to be readily displaced at the carboxyl carbon by a nucleophilic functional group on an affinity chromatography ligand, and x is a positive integer less than or equal to the total number of carboxyl groups in the carboxylated PEI moiety and wherein said R residues are selected from the group consisting of;

##STR4##

2. A 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 pore size of about 50 to 1,000 Angstrom units.

3. A solid phase support of claim 2 wherein R is ##STR5##

4. A solid phase support of claim 1 wherein R is ##STR6##