Bio-rad Laboratories, Inc.

Rensselaer Polytechnic Institute

Russell Frost

Jiali Liao

Hong Chen

Lee Olech

Xuemei He

Steven M Cramer

James Woo

A method for manufacturing a mixed-mode chromatography medium is provided. The method can include, for example, oxidizing diol groups on diol-functionalized solid particles having pores of a median diameter of 0.5 micron or greater with substantially no pores of 0.1 micron or less in diameter and having a diol density of from about 200 to about 300 &#x3bc;mol/mL to aldehyde groups, thereby converting said diol-functionalized solid particles to aldehyde-functionalized solid particles; and coupling amine-functionalized ligands to said aldehyde-functionalized solid particles, said amine-functionalized ligands comprising an amine-substituted hydrophobic group joined to an acid moiety selected from the group consisting of a carboxyl group and a sulfo group.

Solid phase for mixed-mode chromatographic purification of proteins

Proteins are purified by a mixed-mode chromatography system formed by attaching a ligand comprising benzamidoacetic acid to a large-pore support matrix, the only linkage between the ligand and the support matrix being a chain having a backbone of no more than three atoms between a phenyl ring and the support matrix.

Solid supports and ligands are provided for purification of biomolecules by mixed-mode anion exchange-hydrophobic chromatography. Compositions can have the formula Support-(X)&#x2014;N(R1, R2)-R3-L-Ar, or a salt thereof, wherein: Support is a chromatographic solid support; X is a spacer or absent; R1 and R2 are each selected from hydrogen and an alkyl comprising 1-6 carbons; R3 is an alkyl comprising 1-6 carbons or a cyclo alkyl comprising 1-6 carbons; L is NR4, O, or S; wherein R4 is hydrogen or an alkyl comprising 1-6 carbons; and Ar is an aryl. Methods are also provided for using solid supports and ligands to purify biomolecules such as monomeric antibodies.

Anionic exchange-hydrophobic mixed mode

The present disclosure is directed to mixed mode chromatography media comprising a ligand directly attached to a solid support. In some aspects, the ligand has a chemical formula of The mixed mode chromatography media is useful for binding and purifying proteins from a solution.

Mixed mode ligands

Solid supports and ligands are provided for purification of biomolecules by mixed-mode anion exchange-hydrophobic chromatography. Compositions can have the formula Support-(X)&#x2014;N(R, R)&#x2014;R-L-Ar, or a salt thereof, wherein: Support is a chromatographic solid support; X is a spacer or absent; Rand Rare each selected from hydrogen and an alkyl comprising 1-6 carbons; Ris an alkyl comprising 1-6 carbons or a cyclo alkyl comprising 1-6 carbons; L is NR, O, or S; wherein Ris hydrogen or an alkyl comprising 1-6 carbons; and Ar is an aryl. Methods are also provided for using solid supports and ligands to purify biomolecules such as monomeric antibodies.

Proteins are purified by a mixed-mode chromatography system formed by attaching a ligand with cation exchange and hydrophobic functionalities to a large-pore support matrix, the only linkage between the ligand and the support matrix being a chain having a backbone of no more than three atoms between the hydrophobic group and the support matrix.

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Concord, CA, United States of America

Russell Frost