High resolution protein A chromatography employing a chaotropic agent and pH gradient or pH step elution buffer results in improved peak resolution between closely related molecular species. Bispecific antibodies containing a protein A-binding-ablating substitution CH3 domain paired with a protein A-binding CH3 domain are separated with high peak resolution from monospecific antibodies containing a protein A-binding-ablating substituted CH3 domain paired with the protein A-binding-ablating substituted CH3 domain and monospecific antibodies containing a protein A-binding CH3 domain paired with the protein A-binding CH3 domain. Useful chaotropic agents include magnesium chloride and calcium chloride.

The present invention is within the field of chromatography. More precisely, it relates to a novel chromatography medium, namely a hydrophobic medium provided with different lids excluding molecules over a certain size due to the porosity of the hydrophobic medium and/or the porosity of the lid. The invention also relates to use of the separation medium for purification of large molecules, which do not enter the separation medium, as well as small molecules, which enter the separation medium and are eluted from there.

The present invention is within the field of chromatography. More precisely, it relates to a novel chromatography medium, namely a hydrophobic medium provided with different lids excluding molecules over a certain size due to the porosity of the hydrophobic medium and/or the porosity of the lid. The invention also relates to use of the separation medium for purification of large molecules, which do not enter the separation medium, as well as small molecules, which enter the separation medium and are eluted from there.

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 ##STR00001##
The mixed mode chromatography media is useful for binding and purifying proteins from a solution.

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 ##STR00001##
The mixed mode chromatography media is useful for binding and purifying proteins from a solution.

A packing material for liquid chromatography, including particles of a copolymer having a monomer unit derived from a (meth)acrylic acid ester and a monomer unit derived from divinylbenzene, wherein a ratio between the monomer unit derived from a (meth)acrylic acid ester and the monomer unit derived from divinylbenzene is 70 mass % to 90 mass %:30 mass % to 10 mass %, wherein the particles each have a sulfo group bonded to a surface thereof and a structure in which the sulfo group is bonded includes a structure represented by the formula (1):

##STR00001##

where R X and n are as defined herein, and wherein the particles each include the sulfo group at from 40 mol/g to 300 mol/g. Also disclosed is a method of producing the packing material, a column packed with the packing material, and a method of analyzing glycated hemoglobin.

Novel compositions for removing impurities such as, protein aggregates, from a sample containing a protein of interest, e.g., an antibody. Such compositions can be used prior to the virus filtration step during protein purification, to remove aggregates and protect the virus filter from fouling, therefore improving virus filter capacity. A porous solid support including a co-polymer having at least two monomers, wherein at least one of the monomers comprises acrylamide and at least a second monomer comprises a hydrophobic binding group, where the solid support selectively binds protein aggregates, thereby to separate the monomeric protein of interest from the protein aggregates. The method can be performed under neutral to high pH and high conductivity conditions.

Novel compositions for removing impurities such as, protein aggregates, from a sample containing a protein of interest, e.g., an antibody. Such compositions can be used prior to the virus filtration step during protein purification, to remove aggregates and protect the virus filter from fouling, therefore improving virus filter capacity. A porous solid support including a co-polymer having at least two monomers, wherein at least one of the monomers comprises acrylamide and at least a second monomer comprises a hydrophobic binding group, where the solid support selectively binds protein aggregates, thereby to separate the monomeric protein of interest from the protein aggregates. The method can be performed under neutral to high pH and high conductivity conditions.

Described herein is a method of adjusting the composition of a chromatography product to achieve a target enriched proportion of a desired component from an input feed having a lower proportion of the desired component using simulated moving bed (SMB) chromatography wherein the eluent for the SMB apparatus may comprise the very input feed being enriched. The method is exemplified by enriching a high fructose corn syrup from a 42% fructose syrup to a 55% fructose syrup without substantially reducing the dissolved solids concentration of the 55% syrup relative to the input 42% syrup. The 42% syrup is also used as the eluent for the SMB apparatus and may be reconstituted from the raffinate stream by passing the raffinate stream over a glucose isomerase column alone or in combination with a dextrose feed. The method reduces water usage and saves energy by minimizing the need for evaporation to obtain a 55% fructose syrup with a high dissolved solids content.

Described herein is a method of adjusting the composition of a chromatography product to achieve a target enriched proportion of a desired component from an input feed having a lower proportion of the desired component using simulated moving bed (SMB) chromatography wherein the eluent for the SMB apparatus may comprise the very input feed being enriched. The method is exemplified by enriching a high fructose corn syrup from a 42% fructose syrup to a 55% fructose syrup without substantially reducing the dissolved solids concentration of the 55% syrup relative to the input 42% syrup. The 42% syrup is also used as the eluent for the SMB apparatus and may be reconstituted from the raffinate stream by passing the raffinate stream over a glucose isomerase column alone or in combination with a dextrose feed. The method reduces water usage and saves energy by minimizing the need for evaporation to obtain a 55% fructose syrup with a high dissolved solids content.