Abstract: Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Abstract: Provided herein are optimized methods for concentrating large volumes of antibody feedstocks to generate concentrated drug substances by ultrafiltration in a batch-like mode using a fed-batch setup.

Abstract: The disclosure provides a method of separating a protein product, e.g., from a contaminant in a mixture, comprising contacting the mixture with a polishing chromatography matrix. In some embodiments, the polishing chromatography matrix is used together with a retained gradient. In certain embodiments, the retained gradient comprises a pH gradient. In certain embodiments, the mixture comprises a monoclonal antibody product of a prior protein A chromatography.

Abstract: In certain embodiments, the present invention provides a method of measuring the level of hydrophobicity of a chromatographic resin. In certain embodiments, the present invention provides a method of selecting a chromatographic resin condition for purifying a protein of interest from a mixture, wherein the protein of interest has low or no aggregation formation during chromatography. In certain embodiments, the present invention provides a method of selecting a chromatographic resin from a plurality of chromatographic resins for purifying a protein of interest from a mixture, wherein the protein of interest has low or no aggregation formation during chromatography.

Abstract: This disclosure provides a novel method of purifying PEGylated proteins using ion exchange chromatography by loading PEGylated protein having a high concentration, e.g., at least 6 grams/liter on an ion exchange chromatography matrix, and collecting the PEGylated protein.

Abstract: Methods for the production of high purity recombinant protein such as monoclonal antibodies (mAb) using disulfide bond re-oxidation are provided. In particular, the present disclosure provides methods for converting partial molecules (e.g., antibody fragments) to full molecules (e.g., full antibodies) comprising admixing a starting solution comprising the partial molecules with a redox buffer comprising a redox pair which comprises at least one thiol reducing agent (e.g., cysteine) and at least one thiol oxidizing agent (e.g., cystine), wherein the redox buffer re-oxidizes the partial molecules to full molecules. The disclosed methods can be used, e.g., to prevent or mitigate the formation of partial molecules during protein purification, or to reprocess or rescue a solution comprising partial molecules (e.g., a partially degraded pharmaceutical formulation).

Abstract: Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Abstract: This disclosure provides a method of purifying proteins using a combined AEX-CEX chromatography in flow-through mode under an operating condition wherein no automation, engineering control, and/or in-line adjustment is used between the AEX and the CEX. The AEX and CEX (or CEX and AEX) can be connected directly.

Abstract: The present disclosure relates to the use of host cell protein biomarkers to assess disulfide bond reduction in compositions comprising a protein of interest. In some embodiments, the disclosure relates to methods of predicting the occurrence of disulfide bond reduction or low molecular weight protein species in compositions comprising a protein of interest, wherein the expression or activity level of at least one host cell protein is measured and provides a benchmark value associated with the occurrence of disulfide bond reduction or low molecular weight species of said protein of interest. In some embodiments, the disclosure relates to methods of producing a protein of interest, wherein host cells capable of producing the protein of interest are cultured, the expression or activity level of at least one host cell protein is measured, and downstream isolation of the protein of interest is informed by the host cell protein measurements.

Abstract: Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Abstract: Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Abstract: Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Abstract: The present invention is a process for separating a target protein (such as a recombinant protein produced in a cell culture) from a mixture containing the target protein and contaminants (such as cell culture contaminants), by contacting the mixture with a hydrophobic adsorbent comprising branched hydrocarbon functional groups in an aqueous salt solution and collecting the unbound flow-through fraction containing the target protein. In one embodiment, the hydrophobic adsorbent may be a branched alkyl functional group. In another embodiment, the branched alkyl functional group has from 3 to 8 carbon atoms. In another embodiment, the branched alkyl functional group is a tertiary carbon atom, such as tert-butyl.

Abstract: The invention provides methods of controlling pH variations during chromatography. In particular embodiments, the invention includes equilibrating and eluting proteins from chromatography resins have a solid phase that possesses a charged functional group on its backbone under controlled conditions of osmolarity and pH. The invention is particularly useful in the purification of proteins that are sensitive to pH transitions such as, for example, the IL-1 Receptor type II protein.

Abstract: The present invention is a process for separating a target protein (such as a recombinant protein produced in a cell culture) from a mixture containing the target protein and contaminants (such as cell culture contaminants), by contacting the mixture with a hydrophobic adsorbent comprising branched hydrocarbon functional groups in an aqueous salt solution and collecting the unbound flow-through fraction containing the target protein. In one embodiment, the hydrophobic adsorbent may be a branched alkyl functional group. In another embodiment, the branched alkyl functional group has from 3 to 8 carbon atoms. In another embodiment, the branched alkyl functional group is a tertiary carbon atom, such as tert-butyl.

Abstract: The invention provides methods of controlling pH variations during chromatography. In particular embodiments, the invention includes equilibrating and eluting proteins from chromatography resins have a solid phase that possesses a charged functional group on its backbone under controlled conditions of osmolarity and pH. The invention is particularly useful in the purification of proteins that are sensitive to pH transitions such as, for example, the IL-1 Receptor type II protein.