The present invention relates to a method of purifying a target component from a full plasma sample. The present invention further relates to a method of removing one or more target components from a full plasma sample.

The invention relates to poly-amide bonded hydrophilic interaction chromatography (HILIC) stationary phases and novel HILIC methods for use in the characterization of large biological molecules modified with polar groups, known to those skilled in the art as glycans. The invention particularly provides novel, poly-amide bonded materials designed for efficient separation of large biomolecules, e.g. materials having a large percentage of larger pores (i.e. wide pores). Furthermore, the invention advantageously provides novel HILIC methods that can be used in combination with the stationary phase materials described herein to effectively separate protein and peptide glycoforms by eliminating previously unsolved problems, such as on-column aggregation of protein samples, low sensitivity of chromatographic detection of the glycan moieties, and low resolution of peaks due to restricted pore diffusion and long intra/inter-particle diffusion distances.

Disclosed herein are embodiments of methods for oligonucleotide analysis using a novel solid-phase extraction and hydrophilic-interaction liquid chromatography. The unique polar-based retention methods provided herein provide a high-recovery extraction. The methods improve assay reliability and reproducibility and reach picomolar sensitivity with the demonstrably beneficial accurate mass platform. Also disclosed herein are systems and computer program products for performing these methods.

A chromatographic composition includes a solid phase substrate and an ionically-modified hydrophilic ligand coupled to the solid phase substrate. The ionically-modified hydrophilic ligand includes a hydrophilic ligand portion covalently bonded to the solid phase substrate with the hydrophilic ligand including a polar group and a plurality of hydroxyl groups. The ionically-modified hydrophilic ligand also includes an ionic group directly or indirectly coupled to the hydrophilic ligand portion. Methods of producing the chromatographic composition are also provided.

Systems and methods for characterizing low molecular weight (LMW) protein drug product impurities are provided. One embodiment uses hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry analysis. After removal of the N-linked glycans from the protein drug product, for example an antibody drug product, the elution of LMW impurities from the HILIC column was determined by the size of the molecular weight species. In some embodiments, the HILIC separation is performed under denaturing conditions, making the detection of LMW forms using this method highly comparable to both SDS-PAGE and CE-SDS methods. LMW drug product impurities include, but are not limited to light chain, half antibody, H2L, H2, HL, HC, peptide backbone-truncated species, and combinations thereof.

Embodiments of the present disclosure are directed to methods for preparing a target polypeptide from a mixture including the target polypeptide. The method may include contacting the mixture to a hydrophobic interaction chromatography (HIC) apparatus including multiple chromatographic zones. The method may further include passing the target polypeptide through the outlets of at least a first zone and a second zone of the HIC apparatus. A residence time for the mixture including the target polypeptide in a first zone may be approximately the same as a residence time of one or more mobile phases in the second zone.

A filter unit includes a body portion that receives a water filter. An engaging portion is coupled to the body portion. A bypass actuator rotates relative to the engaging portion to rotationally and axially operate a bypass valve of a fluid manifold. The bypass actuator drives axial engagement of the engaging portion with a valve assembly of the fluid manifold. The bypass actuator extends through the body portion and the engaging portion.

A filter unit includes a body portion that receives a water filter. An engaging portion is coupled to the body portion. A bypass actuator rotates relative to the engaging portion to rotationally and axially operate a bypass valve of a fluid manifold. The bypass actuator drives axial engagement of the engaging portion with a valve assembly of the fluid manifold. The bypass actuator extends through the body portion and the engaging portion.

The present disclosure provides chromatography compositions and methods for separation of biomolecules, in particular, stationary phases and chromatography columns for improved separation and resolution of DAR species in antibody drug conjugates (ADCs). In one example, a method comprises: (a) exposing the sample to a chromatographic stationary phase of a reverse phase chromatography column, and (b) applying an eluent to the chromatography column to elute the DAR species of antibody-drug conjugate in their native state, wherein the chromatographic stationary phase comprises: a solid support; and a polymer coating on a surface of the solid support; wherein the polymer coating comprises a copolymer comprising a first hydrophilic monomer and a second hydrophilic monomer, wherein the first hydrophilic monomer and second hydrophilic monomer have different hydrophilicity, wherein the copolymer has a tuned hydrophilicity determined by a molar ratio of the first hydrophilic monomer to the second hydrophilic monomer.