The present disclosure is directed to methods for performing size exclusion chromatographic (SEC) separations. Embodiments of the present disclosure feature methods for improved separations of biomolecule analytes, such as CRISPR-related proteins, nucleotides, and ribonucleoprotein complexes, in SEC, for example, by using hydroxy-terminated polyethylene glycol surface modified stationary phase materials and/or C2/PEG surface modified column hardware.

The present invention provides the use of charged surface reversed phase chromatographic materials along with standard reversed-phase LC and mass spectrometry compatible conditions for the retention, separation, purification, and characterization of acidic, polar molecules, including, but not limited to, organic acids, α-amino acids, phosphate sugars, nucleotides, other acidic, polar biologically relevant molecules. The chromatographic materials of the invention are high purity chromatographic materials comprising a chromatographic surface wherein the chromatographic surface comprises a hydrophobic surface group and one or more ionizable modifier.

The present invention provides the use of charged surface reversed phase chromatographic materials along with standard reversed-phase LC and mass spectrometry compatible conditions for the retention, separation, purification, and characterization of acidic, polar molecules, including, but not limited to, organic acids, α-amino acids, phosphate sugars, nucleotides, other acidic, polar biologically relevant molecules. The chromatographic materials of the invention are high purity chromatographic materials comprising a chromatographic surface wherein the chromatographic surface comprises a hydrophobic surface group and one or more ionizable modifier.

A solid-phase chelator material usable for the purification of proteins. The solid-phase chelator material comprises a solid phase, polyamine groups bound to the solid phase and chelating groups bound to the polyamine groups. At least a part of the polyamine groups is connected with at least two chelating groups per polyamine group. Each chelating group comprises one or several aminopolycarboxylic acid groups (APA groups), with the proviso that the number of APA groups per polyamine group connected with at least two cheating groups is at least three.

Provided is a guard column including a filling part having a length of 2.0 cm to 3.5 cm formed of a filler, in which the filler is made of porous silica gel having a hydrophilized surface and an average particle size of 1.5 μm to 2.5 μm, and a pressure difference when an aqueous solution is fed at a linear flow rate of 2.1 cm/min is 4.0 MPa or more.

A composition includes a hybrid ligand. The hybrid ligand includes an amine group, an amide group or a sulfonamide group, and hydroxyl groups. A first method includes providing a solution containing a first polar analyte and a second polar analyte, applying the solution to a stationary phase including an immobilized hybrid ligand, applying an elution solvent to the stationary phase such that the first polar analyte and the second polar analyte pass through the stationary phase with different elution times, and collecting the first polar analyte at a first elution time and collecting the second polar analyte at a second elution time after the first elution time. A device of a packed column, a cartridge, a tube, a well plate, a membrane, or a planar thin-layer chromatography plate includes a solid support and a hybrid ligand coupled to the solid support. A second method forms an immobilized hybrid ligand.

A composition includes a hybrid ligand. The hybrid ligand includes an amine group, an amide group or a sulfonamide group, and hydroxyl groups. A first method includes providing a solution containing a first polar analyte and a second polar analyte, applying the solution to a stationary phase including an immobilized hybrid ligand, applying an elution solvent to the stationary phase such that the first polar analyte and the second polar analyte pass through the stationary phase with different elution times, and collecting the first polar analyte at a first elution time and collecting the second polar analyte at a second elution time after the first elution time. A device of a packed column, a cartridge, a tube, a well plate, a membrane, or a planar thin-layer chromatography plate includes a solid support and a hybrid ligand coupled to the solid support. A second method forms an immobilized hybrid ligand.

The present disclosure discusses a method of separating and/or purifying polynucleotides. The method includes injecting a sample into a chromatographic column that is packed with a porous sorbent having a pore size that substantially excludes the polynucleotides from the sorbent. This restricted access to the sorbent allows separation of large polynucleotides from each other and from smaller molecular weight impurities.

The present disclosure discusses a method of separating and/or purifying polynucleotides. The method includes injecting a sample into a chromatographic column that is packed with a porous sorbent having a pore size that substantially excludes the polynucleotides from the sorbent. This restricted access to the sorbent allows separation of large polynucleotides from each other and from smaller molecular weight impurities.

The invention relates to a stationary phase medium for adsorption chromatography, which is in form of porous particles suitable for being packed into a chromatographic column. The porous particles are made of cross-linked polymeric material and formed with interconnected macropores to constitute a porous network, through which a mobile phase fluid may flow in a convective manner. The porous particles are substantially free of diffusive pores and, thus, the mass transfer through the porous network is governed by convection alone. The porous particles are fabricated to have irregular granular configurations with rough outer surfaces, so that the convective flow between the porous particles will not be impeded during chromatography process.