Abstract: There is provided a method of operating a liquid chromatography arrangement, the liquid chromatography arrangement comprising: a solvent pump arranged to flow a liquid solvent over a liquid chromatography column; a restrictor arranged to restrict the liquid solvent from leaving the liquid chromatography column; and a liquid pump arranged to provide liquid flow between the liquid chromatography column and the restrictor, the method comprising: flowing the liquid solvent through the liquid chromatography column using the solvent pump; and controlling a liquid pressure within the liquid chromatography column by providing a liquid flow between the liquid chromatography column and the restrictor using the liquid pump.

Abstract: The present disclosure provides compositions and methods for sample processing, particularly for oligonucleotide analysis e.g. analysis of formulated nucleic acid drugs. A composition for pretreating at least one target nucleic acid in a biological mixture provided herein includes a chaotropic agent selected from a substituted guanidine, a substituted amidine, a substituted quaternary amine, or a combination thereof, an optional protease, and/or an optional disulfide-reducing agent. Methods of analyzing at least one target nucleic acid in a biological mixture is also provided herein. Furthermore, the present disclosure provides methods for quantifying at least one target cationic lipid interacting with a nucleic acid.

Abstract: The present invention relates to a method for separating a sample comprising a nucleic acid, the method comprising: (a) loading a sample comprising a nucleic acid onto an anion-exchange column; and (b) eluting the sample from the anion-exchange column using a salt gradient, wherein a mobile phase used in the eluting step comprises a mild ion-pairing cation, and wherein a column temperature of the anion-exchange column is greater than 30° C. The present invention also relates to a method for determining one or more calorimetric properties of a nucleic acid using anion-exchange chromatography.

Abstract: Provided herein chiral derivatization reagents for use in separating and detecting of amine containing enantiomers. The said chiral derivatization reagents provide a combination of improved detectable properties to facilitate various downstream analyses. In particular, the chiral derivatization reagents include at least one chiral carbon atom; at least one strongly basic moiety; at least one chromophore moiety or at least one fluorophore moiety; and at least one reactive group. The present disclosure further provides methods for analyzing amine-containing enantiomeric isomers using a chromatographic separation device and a mass spectroscopy.

Abstract: Methods for quenching a nuclease digestion of a target nucleic acid prior to downstream analysis of the target nucleic acid are disclosed herein. Particularly, methods for controlling the end point of a nuclease digestion prior to sequence analysis of a target nucleic acid is provided. Quenching of a nuclease digestion in the present disclosure employs at least one non-ionic or anionic denaturant combined with an optional reducing agent. The methods presented in this disclosure aids preserving the sample comprising the target nucleic acid or fragments thereof for long term storage and ensures that the effect of contaminating nucleases is eliminated during pretreatment step.

Abstract: The present disclosure relates to methods, compositions and kits useful for the enhanced pH gradient cation exchange chromatography of a variety of analytes. In various aspects, the present disclosure pertains to chromatographic elution buffer solutions that comprise a first buffer salt, a second buffer salt, a third buffer salt, and fourth buffer salt. The first buffer salt may be, for example, a diprotic acid buffer salt, the second buffer salt may be, for example, a divalent buffer salt with two amine groups, the third buffer salt may be, for example, a monovalent buffer salt comprising a single amine group, and the fourth buffer salt may be, for example, a zwitterionic buffer salt. Moreover, the buffer solution has a pH ranging from 3 to 11.

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: A method of capturing human immunoglobulin Fc domains in a biofluid sample is provided. The method includes providing an affinity capture device. The affinity capture device includes a surface having an aptamer that is at least 80% identical to SEQ ID NO 1 immobilized onto the surface of the affinity capture device. The biofluid sample is diluted with a binding buffer. The binding buffer includes (A) tris(hydroxymethyl)aminomethane (Tris), trimethylamine (TES), 2-ethanesulfonic acid (MES), or 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES); (B) a magnesium cation at a concentration between about 10 ?M to about 20 mM; and (C) a total monovalent cation concentration from 0 to no greater than 100 mM. The human immunoglobulin Fc domains in the biofluid sample are adsorbed to the aptamer with the binding buffer.

Abstract: Novel reagents comprising MS active, fluorescent compounds having an activated functionality for reaction with aldehydes and useful in labeling biomolecules such as glycans and methods of making the same are taught and described.

Abstract: Methods for derivatization of biomolecules including glycans or other biopolymers with one or more fluorescent, MS active compounds by reductive amination or rapid tagging in order to produce derivatized glycan having a pKa >7 and between about 200 ? and about 1000 ? of nonpolar surface area are described.

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 2.50% (v/v) difluoroacetic acid and less than about 100 ppb of any individual impurity, especially metal impurities. A sample including the analyte is injected into the mobile phase. The analyte is separated from the sample.

Abstract: Reagents comprising MS active, fluorescent molecules with an activated functionality for reaction with amines useful in tagging biomolecules such as N-glycans and uses thereof are taught and described. In particular, compounds for use as a reagent for rapid fluorescence tagging of biomolecules and enhanced MS signaling are provided. The compounds may have optical centers and therefore may occur in different enantiomeric and diastereomeric configurations. These MS active, fluorescent compounds may have three functional components: (a) a tertiary amino group or other MS active atom; (b) a highly fluorescent moiety, and (c) a reactive group that rapidly reacts with amines. The reactive group provides rapid tagging of desired bio-molecules. The fluorescent moiety provides the fluorescent signal. The tertiary amino group provides the MS signal.

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: 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.

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 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: 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: Methods for derivatization of biomolecules including glycans or other biopolymers with one or more fluorescent, MS active compounds by reductive amination or rapid tagging in order to produce derivatized glycan having a pKa>7 and between about 200 ?2 and about 1000 ?2 of nonpolar surface area are described.

Abstract: Reagents comprising MS active, fluorescent molecules with an activated functionality for reaction with amines useful in tagging biomolecules such as N-glycans and uses thereof are taught and described. In particular embodiments, the MS active, fluorescent molecules are of the compound of the Formula III or of the Formula IV where R1, R2 and R3 are defined herein.

Abstract: The present disclosure is directed to a coating process for chromatographic surfaces. Embodiments of the present disclosure feature a two-step, vapor-liquid phase organosilane deposition method for creating a hydrophilic, non-ionic surface in a chromatographic system.