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. 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.

Abstract: An immobilized enzymatic reactor can include a wall defining a chamber having an inlet and an outlet; a solid stationary phase covalently linked to an enzyme and disposed within the chamber; and a pressure modulator in fluid communication with the chamber and adapted to support continuous flow of a liquid sample comprising a polymer analyte through the inlet, over the solid stationary phase, and out of the outlet under a pressure between about 2,500 and 35,000 psi. In one example, the solid stationary phase includes inorganic/organic hybrid particles in an ultra performance liquid chromatography system, the enzyme is a protease, and the polymer analyte is a polypeptide. The immobilized enzymatic reactor can prepare an analyte for applications such as for hydrogen deuterium exchange mass spectrometry.

Abstract: A clamp assembly includes a rail configured to receive a first fluidic assembly, and a carriage slidably mounted to the rail and configured to receive a second fluidic assembly. The carriage is operable to establish a first fluid tight seal between the first fluidic assembly and a chromatography column received within the claim assembly, and to establish a second fluid tight seal between the second fluidic assembly and the chromatography column.

Abstract: Novel porous materials comprising nanoparticles, use in chromatographic separations, processes for its preparation, and separations devices containing the chromatographic material are described by the instant invention. In particular, the disclosure describes porous inorganic/organic hybrid particles embedded with nanoparticles selected from oxides or nitrides of the following: silicon carbide, aluminum, diamond, cerium, carbon black, carbon nanotubes, zirconium, barium, cerium, cobalt, copper, europium, gadolinium, iron, nickel, samarium, silicon, silver, titanium, zinc, boron, and mixtures thereof.

Abstract: The present disclosure provides methods and kits for detecting an analyte within a sample by using lateral flow testing. Specifically, the present disclosure relates to use of an analyte specific calibration curve for determining quantity of an analyte within a complex sample, wherein the sample is purified/enriched by using a sample preparation method including an affinity resin. The present disclosure further relates to kits including an optional affinity resin, a lateral flow test, and an algorithm corresponding to the analyte specific calibration curve.

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 relates to a method of characterizing proteins in a sample. The method includes: removing non-ionic surfactant from the sample via denaturing size-exclusion chromatography to form a denatured sample; eluting the denatured sample via liquid chromatography to collect fractions of the sample, wherein the fractions of the sample include a protein fraction; lyophilizing the protein fraction to increase protein concentration; reconstituting the lyophilized protein fraction with a buffer comprising a surfactant to denature the protein; digesting the denatured protein fraction with an enzyme; and analyzing the digested protein fraction.

Abstract: The present disclosure relates to protein arrays and methods of using the same for the detection, quantification and characterization of biomolecules that specifically bind to the array among various other biomolecules in a biological sample. Specifically, the present disclosure relates to protein arrays that include a plurality of immunoglobulin molecules derived from shark single-domain heavy chain antibody lacking light-chains but including at least one variable antigen-binding domain with a binding site for an antigen. The immunoglobulin molecules are immobilized on a substrate via a linker. Further encompassed herein are diagnostic devices and kits, comprising the protein array and methods of using same.

Abstract: The present disclosure relates to methodologies, systems, and apparatus for controlling fluid flow within a chromatography system. The chromatography system includes a mobile phase pump configured to pump a liquid mobile phase through a column and a restrictor positioned downstream of the column and upstream of a detector. The system also includes a valve configured to operate in at least two positions. In a first position, the valve is configured to direct the output of the column to bypass the valve and reach the detector, while in the second position the valve directs the output of the column to waste.

Abstract: A method of mass spectrometry is disclosed comprising mass analysing an eluent from a chromatography device and obtaining parent ion data sets and corresponding product ion data sets, and determining whether, in a first product ion data set, one or more product ions are present that are related to one or more parent ions in a corresponding first parent ion data set, based on the mass or mass to charge ratio and/or intensity of the one or more product ions and the one or more parent ions. If it is determined that the one or more product ions are present, the method further comprises removing the one or more product ions from one or more second product ion data sets to produce one or more second modified product ion data sets and/or removing ions other than the one or more product ions from the first product ion data set to produce a first modified product ion data set.

Abstract: The present disclosure relates to an apparatus for regulating the average mobile phase density or pressure in a carbon dioxide based separation system. The apparatus includes a controller, a set of pressure or density sensors and a set of instructions capable of determining the pressure drop across a column and adjusting at least one system component or parameter to achieve a pre-determined average mobile phase density or pressure in the system.

Abstract: A mixed-mode chromatography method for the determination of phosphorylated sugars in a sample is provided. The mixed-mode chromatography method includes obtaining a sample comprising at least one phosphorylated sugar. The sample is introduced onto a chromatography system. The chromatography system includes a column having a stationary phase material contained inside the column. The stationary phase material has a surface comprising a hydrophobic surface group and at least one ionizable modifier. The sample with a mobile phase eluent is flowed through the column, where the at least one phosphorylated sugar is substantially resolved and retained within seven minutes. The mobile phase eluent includes water with an additive and acetonitrile with the additive. The mobile phase eluent has a pH less than 6. The at least one phosphorylated sugar is detected using a detector.

Abstract: The invention provides filters and methods of assembling filters. In an exemplary embodiment, the filter includes a porous element, a compression element, and a housing. The compression element can be configured to receive the porous element, thereby forming an assembly. For example, the compression element can receive the porous element in a slip-fit relationship. The housing can have an opening formed therein that is configured to receive the assembly. In some embodiments, the assembly can be retained within the opening when the assembly is received therein. For example, the opening can receive the assembly in a press-fit relationship.

Abstract: The present disclosure relates to methodologies, systems, apparatus, and kits for controlling fluid flow within a chromatography system. A makeup pump is configured to pump a makeup fluid into the chromatography system downstream of the column. A first restrictor is located upstream of a detector and downstream of both the makeup pump and the column. Decreasing an output volume of the makeup pump can direct an output from the column through the first restrictor to the detector. Increasing an output volume of the makeup pump can direct the output from the column to a second restrictor located downstream of the makeup pump and the column and in parallel with the first restrictor and the detector.

Abstract: Sorbents, devices, kits and methods useful for sample treatment are disclosed herein. In particular embodiments, described are inorganic/organic hybrid sorbent particles comprising (a) a core region that comprises a silica component and (b) a surface region that comprises an organic copolymer comprising at least one hydrophobic organic monomer and at least one hydrophilic organic monomer.

Abstract: 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 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, 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.

Abstract: The present disclosure discusses a method of separating a sample (e.g., B vitamins and their vitamers) 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 provides an inert barrier that is beneficial for B vitamin analysis, including improved peak shape (less tailing and narrower peak), high sensitivity, and no carry-over.

Abstract: A vial cap for a sample vial includes a cylindrical cap body defining a first circular opening at a first end. The vial cap also includes a cap lid extending radially inward from a second end of the cylindrical cap body. The cap lid defines a second circular opening. The vial cap also includes a septum located within the cylindrical cap body and in contact with the cap lid. The septum spans the second circular opening defined by the cap lid portion. The vial cap also includes a number of flexible threads extending from an internal surface of the cylindrical cap body.

Abstract: An error compensation system and method may include applying a mechanical load to a reference sample to obtain a load measurement signal from the load sensor and a displacement measurement signal from the displacement sensor, calculating a transfer function to create a load filter and a displacement filter to be applied to the load measurement signal and the displacement measurement signal, respectively, applying the load filter to the load measurement signal to calculate a load compensation value, and applying the displacement filter to the displacement measurement signal to calculate a displacement compensation value, and determining the compensated value by comparing the load compensation value with the displacement compensation value, wherein the compensated value is determined prior to testing a specimen so that the compensated value is used to automatically correct a measured deflection of the specimen to arrive at an actual specimen deflection.

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.