Abstract: Methods of analyzing glycosylated biomolecules include the steps of producing a deglycosylation mixture of biomolecules deglycosylated by natural or synthetic enzymatic or chemical techniques; providing a reagent solution having a labeling reagent in a polar aprotic, non-nucleophilic organic solvent; and mixing the deglycosylation mixture with the reagent solution in an excess of labeling reagent to produce derivatized glycosylamines. The method steps can be carried out purposefully without depletion of protein matter. A quenching solution can be added to the reaction mixture so that the pH of the reaction mixture is shifted to above 10. The yield of derivatized glycosylamines can be in an amount of about 80 to about 100 mole percent of the reaction mixture with minimal overlabeling, less than 0.2 mole percent.

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 present disclosure is directed to nonporous polymer particles having an average particle size of 1 to 10 micrometers and being functionalized with an immunoglobulin-binding protein. The functionalized particles of the present disclosure can be used to purify a range of immunoglobulins with affinity to the immunoglobulin-binding protein.

Abstract: The present disclosure is directed to nonporous polymer particles having an average particle size of 1 to 10 micrometers and being functionalized with streptavidin. The streptavidin functionalized particles of the present disclosure can be bound to biotinylated antibodies, biotinylated antigen-binding fragments thereof, or biotinylated oligonucleotides to create customizable affinity chromatography columns. That is, the functionalized nonporous polymer particles can be packed in a stainless steel, titanium, or other metal or metal alloy column for affinity high pressure liquid chromatography (HPLC).

Abstract: The present disclosure relates to a filter. The filter includes a porous element, a compression element and a housing. At least a portion of the porous element is coated with an alkylsilyl coating. The compression element is configured to receive the porous element thereby forming an assembly. The housing has an opening formed therein. The opening is configured to receive the assembly. The assembly is retained within the opening when the assembly is received therein.

Abstract: A sample preparation device is provided for enriching a component of a sample. The sample preparation device includes a surface in fluid communication with the sample, an alkylsilyl coating disposed on the surface, and an affinity ligand or an enzyme covalently bonded to the alkylsilyl coating.

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 relates to the determination of analytes in a sample using chromatography. The present disclosure provides methods of separating an analyte from a sample. The method includes introducing a sample comprising the analytes into a chromatographic system. The chromatographic system has a flow path disposed in an interior of the chromatographic system, at least a portion of the flow path having an active coating, and a chromatographic column having a stationary phase material in an interior of the chromatographic column that facilitates separation of the analytes in the sample through interaction with at least one analyte in the sample. The active coating is selected to interact with at least one analyte in the sample through (1) a repulsive force, (2) a secondary interaction, or (3) a retention mechanism distinct from the interaction with the stationary phase material.

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 technology relates to a method for improved recovery of phosphorylated compounds (e.g., phosphorylated glycans). In particular, the present technology utilizes certain chelator additives in a solution to wash and elute the phosphorylated compounds from a solid phase extraction cartridge.

Abstract: A sample preparation device is provided for enriching a component of a sample. The sample preparation device includes a surface in fluid communication with the sample, an alkylsilyl coating disposed on the surface, and an affinity ligand or an enzyme covalently bonded to the alkylsilyl coating.

Abstract: The present technology relates to a method of separating a sample comprising oligonucleotides. The method includes injecting a polyphosphonic acid at a concentration of between about 0.01 M to about 1 M into the sample comprising oligonucleotides. The method also includes flowing the sample and polyphosphonic acid through a liquid chromatography column and separating the oligonucleotides.

Abstract: Stationary phase materials for performing size exclusion chromatography are provided. The stationary phase materials include porous inorganic-organic hybrid particles surface-modified with a hydroxy-terminated polyethylene glycol silane.

Abstract: Provided herein are compositions, kits and methods that may be useful for improving the efficiency of solid phase extraction to separate oligonucleotides from a complex matrix. Particularly, the present technology relates to a sorbent material including porous particles, the surface of which is modified with a ligand that includes one or more bridging alkyl substituted amines and at least two siloxyl terminal groups.

Abstract: The present disclosure pertains to sample preparation devices useful for affinity capture and purification that include one or more internal structures that comprise a reservoir, a well, a fluid passageway, sorbent particles, and a filter element that blocks passage of the affinity sorbent particles, which sample preparation devices combine the attributes of both dispersive and flow through designs into a single sample preparation device. The present disclosure also pertains to kits that contain and methods that use such sample preparation devices.

Abstract: 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. The method includes introducing a sample comprising the analytes into a chromatographic system. The chromatographic system has a flow path disposed in an interior of the chromatographic system, at least a portion of the flow path having an active coating, and a chromatographic column having a stationary phase material in an interior of the chromatographic column that facilitates separation of the analytes in the sample through interaction with at least one analyte in the sample. The active coating is selected to interact with at least one analyte in the sample through (1) a repulsive force, (2) a secondary interaction, or (3) a retention mechanism distinct from the interaction with the stationary phase material.

Abstract: The present technology relates to a method for improved recovery of phosphorylated compounds (e.g., phosphorylated glycans). In particular, the present technology utilizes certain chelator additives in a solution to wash and elute the phosphorylated compounds from a solid phase extraction cartridge.

Abstract: The present disclosure describes methods for analyzing mRNA poly(A) tail sequences with a high resolution to determine poly(A) tail length and heterogeneity. The method involves digesting an mRNA molecule to liberate a poly(A) tail, preparing a chromatographic sample comprising the mRNA poly(A) tails, preparing a second chromatographic sample comprising a reference sequence comprising a mRNA poly(A) tails having a predetermined length, separating the first and second samples by a chromatography method, which result in one or more chromatograms, and determining a sequence length of the mRNA poly(A) tails by comparing the chromatograms of the first and second samples. Chromatography methods for analyzing the poly(A) tail may include ultraviolet size-exclusion chromatography (SEC UV), ultraviolet ion-pair reversed-phase liquid chromatography (IP RP LC UV), ultra high performance liquid chromatography (UPHLC), and combinations thereof.

Abstract: The present disclosure discusses a method of separating and/or purifying polynucleotides. The method includes injecting a sample into cartridge that is packed with a porous sorbent between opposing frits. Physical properties of the cartridge along with the use of hydrophilic fits allows efficient desalting of biopolymers using a vacuum to pull the sample through the cartridge.

Abstract: The present disclosure pertains to non-porous composite particles that are non-porous, polymer-based particles. In various embodiments, a non-porous polymer core is surface modified. In various embodiments, a non-porous hybrid organic-inorganic material is in contact with the modified surface of the core, and a bonding material is in contact with the non-porous hybrid organic-inorganic material. The present disclosure pertains to chromatographic separation devices that comprise such non-porous composite particles.