Affinity membrane and method of preparation
Inventors
ZHOU, JINXIANG • Temples, Graham • Henn, Daniel
Assignees
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Publication Number
US-11918957-B2
Publication Date
2024-03-05
Expiration Date
Abstract
A method for preparing an adsorptive media for binding biologic molecules comprising immersing a macroporous support in a first solution of a coupling reagent in a solvent solution for attachment of said coupling reagent to form coupling groups; and, immersing said macroporous support in an incubating solution selected from the group consisting of ligand, nucleotide, oligonucleotide, peptide, polypeptide, protein, and enzyme solutions having an affinity to a biologic target molecule to couple one of said ligands, nucleotides, oligonucleotides, peptides, polypeptides, proteins, and enzymes to at least a portion of said coupling groups of said macroporous support for binding with said biologic target molecule when exposed to said macroporous support.
Core Innovation
The invention relates to methods for preparing an adsorptive media for binding biologic molecules. A macroporous support is immersed or contacted in a first solution comprising a coupling reagent and a first solvent to form coupling groups on at least a portion of the macroporous support. The coupling groups are then contacted with an incubating solution to couple a molecule having an affinity to a biological target to the coupling groups on the macroporous support.
The incubating solution comprises water and an organic solvent, where the organic solvent is present in an amount tied to the cloud point of the ligand. In some embodiments the organic solvent amount ranges from 90% to 110% of a volumetric amount of the organic solvent at the cloud point of the ligand, while other embodiments define organic solvent amounts ranging from 75% to 125% of the cloud point amount or from 50 vol-% to 99 vol-%.
The affinity molecule coupled to the coupling groups includes a nucleotide, an oligonucleotide, a peptide, a polypeptide, a protein, or any combination thereof. The document also describes preparing adsorptive media with macroporous cellulose membranes and forming coupling groups on such membranes prior to incubating with the affinity molecule in a water/organic-solvent incubating solution.
Claims Coverage
The independent claims include three methods for preparing adsorptive media. They share forming coupling groups on a macroporous support and coupling an affinity molecule in a water/organic-solvent incubating solution, with quantitative constraints on the organic solvent amount relative to ligand cloud point and specified classes of affinity molecules.
Cloud-point-relative organic solvent incubating solution for affinity coupling
Form coupling groups on a macroporous support using a coupling reagent in a first solution, then couple a molecule having an affinity to a biological target to at least a portion of the coupling groups using an incubating solution comprising water and an organic solvent present in an amount ranging from 90% to 110% of a volumetric amount of the organic solvent at the cloud point of the ligand.
Affinity molecule coupled to coupling groups on a macroporous support
Contact a macroporous support with a first solution comprising a coupling agent and a first solvent to form a macroporous support having coupling groups; and contact the macroporous support having coupling groups with an incubating solution comprising water, an organic solvent, and a molecule having an affinity to a biological target to couple the ligand with at least some of the coupling groups.
Organic solvent amount relative to ligand cloud point and specified affinity-molecule classes
Wherein the organic solvent is present in the incubating solution in an amount ranging from 75% to 125% of amount of organic solvent at a cloud point of the ligand; and wherein the molecule comprises a nucleotide, an oligonucleotide, a peptide, a polypeptide, a protein, or any combination thereof.
Macroporous cellulose membrane coupling groups and nucleotide/oligonucleotide coupling
Contact a macroporous cellulose membrane with a first solution comprising a coupling agent and a first solvent to form a macroporous cellulose membrane having coupling groups; and contact the macroporous cellulose membrane having coupling groups with an incubating solution comprising water, an organic solvent, and a molecule having an affinity to a biological target to couple the molecule with at least some of the coupling groups, wherein the organic solvent is present in the incubating solution in an amount ranging from 50 vol-% to 99 vol-% and wherein the molecule comprises a nucleotide, an oligonucleotide, or any combination thereof.
The inventive coverage centers on coupling an affinity molecule to coupling groups formed on a macroporous support, including macroporous cellulose membranes, using an incubating solution containing water plus an organic solvent defined relative to ligand cloud point or within a specified vol-% range. The independent claims cover nucleotide, oligonucleotide, peptide, polypeptide, and protein embodiments, with dependent claims further narrowing supports and quantitative constraints.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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