Bio-rad Laboratories, Inc.

Rensselaer Polytechnic Institute

Jiali Liao

Hong Chen

Christopher Belisle

Xuemei He

Russell Frost

Yueping Xu

Lee Olech

Tom R Berkelman

Edwin Lozano

Steven M Cramer

James W Xu

Haihui Yuan

James Woo

Fang Song

Xiaozhong Bao

Juan You

Wenli Cao

Methods of preparing polymer-filled chromatography resin and their uses are provided.

Method of preparing polymer-filled chromatography resin

Chromatography resins having anionic exchange-hydrophobic mixed mode ligands and methods of using such resins are provided.

Anionic exchange-hydrophobic mixed mode chromatography resins

Chromatography resins having anionic exchange-hydrophobic mixed mode ligands, that are useful for purifying target biomolecules using anionic exchange (i.e., where the ligand is positively charged) and hydrophobic mixed mode chromatography. The chromatography resins allow for efficient purification of target biomolecules (e.g., recombinant proteins, antibodies, antibody-drug conjugates, or antibody derivatives including, but not limited to, antibody fragments and antibody fusions) from a sample, and have been found to be useful in purifying monomeric target biomolecules from aggregate target biomolecules. In an embodiment, the chromatography resins are useful for separating antibodies from one or more components (e.g., contaminants) in the sample.

Anionic exchange-hydrophobic mixed mode chromatography resin

The subject invention pertains to proteins are purified by a mixed-mode chromatography system formed by attaching a ligand with cation exchange and hydrophobic 1,3-dioxoisoindolin-2-yl group functionalities to a large-pore support matrix, the only linkage between the ligand and the support matrix being a chain having a backbone of one, two, three, four, or five atoms between the hydrophobic group and the support matrix.

Mixed mode cation exchange chromatography ligands based on 1,3-dioxoisoindolin-2-yl structures

Chromatography resins having mixed mode ligands and methods of using such resins are provided.

Chromatography resin having an anionic exchange-hydrophobic mixed mode ligand

Methods and kits for purifying protein-nanoparticle conjugates are provided. In some embodiments, a multimodal medium having a size exclusion mode and a capture mode is used to purify the protein-nanoparticle conjugates.

Protein-nanoparticle conjugate purification methods

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

Polymer-filled ceramic apatites and their uses are provided.

Agarose-filled ceramic apatite

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.

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San Ramon, CA, United States of America

Jiali Liao