Methods for identifying viral protein constituents and quantifying the relative abundance of such viral protein constituents in a sample of viral particles are disclosed. In embodiments, the methods include first-dimension chromatography to separate intact viral capsid components of the sample, online denaturation of the viral capsid components to produce intact viral proteins, second-dimension chromatography to separate the viral proteins, and mass spectrometry to determine the masses of the viral proteins and identify the viral protein constituents of the sample.

The present invention provides novel chromatographic materials, e.g., for chromatographic separations, processes for their preparation and separations devices containing the chromatographic materials. The preparation of the inorganic/organic hybrid materials of the invention wherein a surrounding material is condensed on a superficially porous hybrid core material will allow for families of different hybrid packing materials to be prepared from a single core hybrid material. Differences in hydrophobicity, ion-exchange capacity, chemical stability, surface charge or silanol activity of the surrounding material may be used for unique chromatographic separations of small molecules, carbohydrates, antibodies, whole proteins, peptides, and/or DNA.

The present disclosure relates to the determination of analytes in a sample using chromatography. The present disclosure provides methods of separating an analyte from a sample. A mobile phase is flowed through a chromatography column. The mobile phase includes about 0.005% (v/v) to about 0.20% (v/v) difluoroacetic acid and less than about 100 ppb of any individual metal impurity. A sample including the analyte is injected into the mobile phase. The analyte is separated from the sample.

The present invention provides methods to improve the sensitivity of detecting glycosylamines released from glycoconjugates, such as glycoproteins or glycopeptides, by enzymatic digestion when labeling them with amine-reactive dyes.

In some aspects, the present disclosure pertains to sample treatment methods that comprise: contacting an acidic elution solution that is free of primary amine, secondary amine and thiol groups with a sorbent having bound target antibody and separating the elution solution from the sorbent, thereby releasing bound target antibody from the sorbent and forming a first collection fraction that comprises the elution solution and released target antibody; contacting the sorbent with a neutralization buffer solution that is free of primary amine, secondary amine and thiol groups and separating the neutralization buffer solution from the sorbent, thereby forming a second collection fraction that comprises the neutralization buffer solution; and forming a neutralized solution that comprises the first collection fraction and the second collection fraction. In other aspects, the present disclosure pertains to kits for performing such sample treatment methods.

A separating medium for hydrophilic interaction chromatography useful in separating hydrophilic compounds. The hydrophilic interaction chromatography separating medium, which is formed from a support and a ligand carried by the support, is a separating medium wherein the ligand is a (meth)acrylic polymer having a constituent unit derived from the compound indicated by formula (I). ##STR00001##

The invention discloses a separation matrix for purification of biological particles, comprising a plurality of particles having a porous core entity and a porous shell entity covering the core entity, wherein the core entity comprises at least 50 micromole/ml primary amines present on covalently attached ligands displaying at least two primary amines per ligand and the shell entity comprises less than 20 micromole/ml primary amines The invention further discloses a method of purifying biological particles and a method of manufacturing a separation matrix.

In one aspect, the present invention provides a chromatographic stationary phase material for various different modes of chromatography represented by Formula 1: [X](W).sub.a(Q).sub.b(T).sub.c (Formula 1). X can be a high purity chromatographic core composition having a surface comprising a silica core material, metal oxide core material, an inorganic-organic hybrid material or a group of block copolymers thereof. W can be absent and/or can include hydrogen and/or can include a hydroxyl on the surface of X. Q can be a functional group that minimizes retention variation over time (drift) under chromatographic conditions utilizing low water concentrations. T can include one or more hydrophilic, polar, ionizable, and/or charged functional groups that chromatographically interact with the analyte. Additionally, b and c can be positive numbers, with the ratio 0.05≤(b/c)≤100, and a≥0.

Methods for preparing labeled glycosylamines from a complex matrix are provided. The methodology includes the steps of: denaturing glycoproteins in a complex matrix to form a denatured complex matrix mixture; loading the denatured complex matrix mixture onto a MWCO filtration device; adding a glycosidase enzymatic solution onto the MWCO filtration device to form a deglycosylated complex matrix mixture comprising glycosylamines; collecting glycosylamines released from the MWCO filtration device; and derivatizing glycosylamines with a rapid tagging reagent to form a plurality of labeled glycosylamines suitable for detection in various liquid chromatography systems and detectors.

A purification agent which includes a compound having a betaine structure, and which is for a sugar chain having a length equal to or longer than that of a monosaccharide or for a glycopeptide having a sugar chain having a length equal to or longer than that of a monosaccharide.