Abstract: The present disclosure relates to a method of reducing degradation of sample components in a liquid chromatography system. The method utilizes a masked metal frit to prevent or reduce metal surfaces from becoming catalytically active. The masked metal frit is a metal based frit that includes a coating on its exterior surfaces to mask or prevent contact between the organic solvents (and/or any analyte or other related solvent) and the underlying metal. The coating is a vapor deposited inorganic-organic hybrid coating, such as a vapor deposited C2 coating.

Abstract: The present disclosure is directed to methods of characterizing a system containing a chromatographic column. The methods can include introducing a sample comprising a positive control and a negative control to the system containing a chromatographic column, wherein the positive control is a sensitive probe that interacts with the system and the negative control is substantially non-interacting with the system; after passing the sample through the chromatographic column, detecting the positive control and the negative control; and determining system suitability by comparing the amount of detected positive control to negative control. In some embodiments, determining system suitability (e.g., inertness of sample to the system) is accomplished by determining a ratio of detected positive control to negative control.

Abstract: The present disclosure is directed to methods for evaluating system inertness, such as the inertness of a LC or other fluidic system. Some methods are directed to tests wherein the column has been removed prior to injecting a sample including a positive (e.g., metal reacting moiety) control into the system. Some methods can include: (1) repeatedly injecting the sample into a system, the system comprising: fluidic paths wherein interior surfaces of the fluidic paths define wetted surfaces, and wherein at least a portion of the wetted surfaces of the fluidic flow path are coated with an inert coating, wherein the inert coating is inert to at least one analyte in the sample; (2) detecting a value associated with the positive control; and (3) analyzing values associated with the detected positive control to determine system inertness.

Abstract: The present disclosure is directed to stationary phase materials (e.g., porous inorganic-organic hybrid particles) for performing size exclusion chromatography. Embodiments of the present disclosure feature hydroxy-terminated polyethylene glycol surface modified stationary phase materials.

Abstract: The present disclosure is directed to methods for performing size exclusion chromatography. Embodiments of the present disclosure feature methods for improving separations of proteinaceous analytes in size exclusion chromatography, for example, by using low concentrations of amino acids or derivatives thereof in the mobile phase.

Abstract: The present disclosure is directed to stationary phase materials for performing size exclusion chromatography. Embodiments of the present disclosure feature hydroxy-terminated polyethylene glycol surface modified silica particle stationary phase materials, which are optionally also methoxy-terminated polyethylene glycol surface modified.

Abstract: 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.

Abstract: Provided herein are stationary phase compositions comprising a chromatographic surface of porous or non-porous core material comprising a surface modifier for use in chromatographic separations.

Abstract: The present disclosure relates to a kit for sample preparation, the kit including a solid support surface with a polymer coating covering the solid support surface, wherein the polymer coating reduces undesired interactions between the sample and the solid support surface, a buffer comprising arginine and methionine, and a vessel for containing the solid support surface and the buffer.

Abstract: The present disclosure relates to a system for a composition for protein denaturation. The composition includes a non-nucleophilic denaturant comprising a substituted guanidine, wherein the denaturant has a pKa value greater than about 10, and wherein the concentration of the substituted guanidine is less than 250 mM.

Abstract: The present disclosure relates to a system for separating a sample including a device and a positive pressure source. The device can include a housing with a proximal end having an interface and a proximal end opening, a distal end opening opposite the proximal end opening, and an interior wall defining an interior of the housing; a bottom frit connected to the interior wall, extending across the interior of the housing, and located proximately to the distal end to minimize sample loss; and a resin disposed within the interior of the housing between the bottom frit and the proximal end. The positive pressure source can connect to the interface of the proximal end to apply positive pressure to the sample. A controller can control the applied positive pressure to the sample via the positive pressure source according to a relationship between the bottom frit, the resin, and the positive pressure.

Abstract: In various aspects, the present disclosure pertains to materials (e.g., kits, column assemblies, liquid chromatography systems, etc.) methods for performing liquid chromatography that employ a first column (e.g., a trapping column) and a second column (e.g., an analytical column). The first column comprises a first chromatographic material having a first chromatographic surface that comprises first hydrophobic surface groups and first ionizable surface groups having a first pKa value. The second column comprises a second chromatographic material having a second chromatographic surface that comprises second hydrophobic surface groups and (a) permanently ionized surface groups or (b) second ionizable surface groups having a second pKa value. The first hydrophobic surface groups have a hydrophobicity that is less than a hydrophobicity of the second hydrophobic surface groups.

Abstract: The present disclosure relates to the use of vapor deposition coated flow paths for improved chromatography and sample analysis using liquid chromatography-mass spectrometry (LC/MS) or liquid chromatography-optical detection (LC/UV). More specifically, this technology relates to separating and quantitation of analytes (e.g., phospho prodrugs and its phosphorylated metabolites) from a sample matrix (e.g., mammalian blood, plasma) using chromatographic devices and fluidic systems having coated flow paths. The LC-MS or LC-UV techniques provide improved recovery, peak shape and dynamic range in the analysis of the prodrug and its metabolites.

Abstract: The present disclosure relates to a hetero-functional coating applied on a solid support. The coating includes a first functionality for conjugating biomolecules for the analysis of a protein or nucleic acid, and a second functionality for preventing undesired interactions between analytes of interest and the surface of solid support.

Abstract: The present disclosure relates to a method of digesting a sample comprising protein, the method comprising: adding the sample to an apparatus containing a buffer and a solid support surface comprising a surface coating, wherein the surface coating immobilizes enzyme while reducing undesired interactions between the sample and the solid support surface; immobilizing enzymes on the surface coating for digestion of the protein of the sample; and heating the sample to complete a heat-assisted digestion of the protein.

Abstract: 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.

Abstract: The present disclosure provides devices, systems, kits and methods useful for quantitation of biomolecules such as intact proteins and nucleic acids.

Abstract: The present disclosure discusses a method of separating a sample (e.g., peptide compound) including coating a flow path of a chromatographic system; injecting the sample into the chromatographic system; flowing the sample through the chromatographic system; separating the sample; and analyzing the separated sample. In some examples, the coating applied to the surfaces defining the flow path is non-binding with respect to the sample—and the separated sample. Consequently, the sample does not bind to the low-binding surface of the coating (e.g., organosilica coating) of the flow path. The applied coating can reduce peak tailing and increase analyte recovery for the sample of the chromatographic system.

Abstract: 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.

Abstract: The present technology relates to a customizable sample preparation device (e.g., liquid sample preparation device, such as a purification, clean-up, or separation device). In particular, the present technology relates to customizable devices formed from modular segments tailored to address one or more of the following: to optimize different sample and elution volumes, to incorporate various connection mechanisms to liquid handlers, to incorporate various liquid dispensing flow conditions, and to fulfill broad applications through the selection of specific resin for sample preparation. Some embodiments are directed to affinity capture devices, such as for example, Protein A affinity capture devices, utilizing a polymethacrylate-based resin. Other resins as well as other materials for processing a liquid sample are described herein.