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.

Abstract: This present disclosure relates to engineered protease enzymes, including trypsin, Lys-C and Asp-N proteases, that have enhanced autolysis resistance. Also disclosed herein are methods of using such engineered enzymes for improving detection of target analyte proteins in an analytical assay.

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: The present disclosure describes methods, kits, and systems for digesting polyribonucleotides. The method involves selectively forming oligonucleotide (e.g., DNA:RNA or RNA:RNA) duplexes with single-stranded target RNA and then using sequence-specific nucleases that only act on RNA within duplexes to selectively cleave the target RNA into smaller fragments. Additional sequence-specific ribonucleases may be used to provide additional cuts of the target RNA at predetermined sites. By forming duplexes to increase the availability of nucleases that may be applied to cleave the single-stranded target RNA and selectively control where the target RNA is cleaved, the target RNA may be digested into fragments within controllable size ranges that are optimal for polynucletide analysis, such as by liquid chromatography and mass spectrometry.

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 methods of sample preparation from lipid nanoparticle (LNP)-encapsulated nucleic acids utilizing solid-phase extraction techniques.

Abstract: A device for processing samples is disclosed. Interior surfaces of the device, which come in contact with fluids, define wetted surfaces. A portion of the wetted surfaces are coated with an alkylsilyl coating having the Formula I: R1, R2, R3, R4, R5, and R6 are each independently selected from (C1-C6)alkoxy, —NH(C1-C6)alkyl, —N((C1-C6)alkyl)2, OH, ORA, and halo. RA represents a point of attachment to the interior surfaces of the fluidic system. At least one of R1, R2, R3, R4, R5, and R6 is ORA. X is (C1-C20)alkyl, —O[(CH2)2O]1-20-, —(C1-C10)[NH(CO)NH(C1-C10)]1-20-, or —(C1-C10)[alkylphenyl(C1-C10)alkyl]1-20-.

Abstract: A device for separating analytes is disclosed. The device has a sample injector, sample injection needle, sample reservoir container in communication with the sample injector, chromatography column downstream of the sample injector, and fluid conduits connecting the sample injector and the column. The interior surfaces of the fluid conduits, sample injector, sample reservoir container, and column form a flow path having wetted surfaces. A portion of the wetted surfaces of the flow path are coated with an alkylsilyl coating that is inert to at least one of the analytes. The alkylsilyl coating has the Formula I: R1, R2, R3, R4, R5, and R6 are each independently selected from (C1-C6)alkoxy, —NH(C1-C6)alkyl, —N((C1-C6)alkyl)2, OH, ORA, and halo. RA represents a point of attachment to the interior surfaces of the fluidic system. At least one of R1, R2, R3, R4, R5, and R6 is ORA. X is (C1-C20)alkyl, —O[(CH2)2O]1-20, -(C1-C10)[NH(CO)NH(C1-C10)]1-20-, or -(C1-C10)[alkylphenyl(C1-C10)alkyl]1-20-.

Abstract: The present disclosure is directed to liquid chromatography columns and methods utilizing a stationary phase sorbent having two or more gradient factors. Each of the two or more gradient factors varies in a progressive manner along a length of the stationary phase sorbent in a direction from an inlet to an outlet of the column (i.e., along a length of the column). As a result of including these new continuous stationary phase gradients, new selectivities allowing for separation and analysis of complex samples including large biomolecules is achievable.

Abstract: The present disclosure discusses a method of separating and/or purifying polynucleotides. The method includes injecting a sample into a chromatographic column that is packed with a porous sorbent having a pore size that substantially excludes the polynucleotides from the sorbent. This restricted access to the sorbent allows separation of large polynucleotides from each other and from smaller molecular weight impurities.

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 µM 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: Methods are provided for making rapid labeled dextran ladders and other calibrants useful in liquid chromatography. The methodologies include a two-step process comprising a reductive amination step of providing a reducing glycan and reacting it with a compound having a primary amine to produce an intermediate compound. The intermediate compound is then rapidly tagged with a rapid tagging reagent to produce the rapid labeled dextran ladder.

Abstract: The present disclosure is directed to methods for performing size exclusion chromatographic (SEC) separations. Embodiments of the present disclosure feature methods for improved separations of biomolecule analytes, such as CRISPR-related proteins, nucleotides, and ribonucleoprotein complexes, in SEC, for example, by using hydroxy-terminated polyethylene glycol surface modified stationary phase materials and/or C2/PEG surface modified column hardware.

Abstract: The present invention relates to a method for analyzing a sample, the method comprising: (a) incubating a sample comprising an analyte with at least one enzyme to produce a digestion mixture comprising fragments of the analyte; (b) loading the digestion mixture onto a reversed-phase chromatography column; and (c) performing reversed-phase chromatography on the digestion mixture, wherein the at least one enzyme is hydrophobically modified to increase a retention time of the at least one enzyme such that the at least one enzyme elutes from the reversed-phase chromatography column later than the fragments of the analyte.

Abstract: The present invention provides the use of charged surface reversed phase chromatographic materials along with standard reversed-phase LC and mass spectrometry compatible conditions for the retention, separation, purification, and characterization of acidic, polar molecules, including, but not limited to, organic acids, ?-amino acids, phosphate sugars, nucleotides, other acidic, polar biologically relevant molecules. The chromatographic materials of the invention are high purity chromatographic materials comprising a chromatographic surface wherein the chromatographic surface comprises a hydrophobic surface group and one or more ionizable modifier.

Abstract: A device for processing samples is disclosed. Interior surfaces of the device, which come in contact with fluids, define wetted surfaces. A portion of the wetted surfaces are coated with an alkylsilyl coating having the Formula I: R1, R2, R3, R4, R5, and R6 are each independently selected from (C1-C6)alkoxy, —NH(C1-C6)alkyl, —N((C1-C6)alkyl)2, OH, ORA, and halo. RA represents a point of attachment to the interior surfaces of the fluidic system. At least one of R1, R2, R3, R4, R5, and R6 is ORA. X is (C1-C20)alkyl, —O[(CH2)2O]1-20—, —(C1-C10)[NH(CO)NH(C1-C10)]1-20—, or —(C1-C10)[alkylphenyl(C1-C10)alkyl]1-20—.

Abstract: A device for separating analytes is disclosed. The device has a sample injector, sample injection needle, sample reservoir container in communication with the sample injector, chromatography column downstream of the sample injector, and fluid conduits connecting the sample injector and the column. The interior surfaces of the fluid conduits, sample injector, sample reservoir container, and column form a flow path having wetted surfaces. A portion of the wetted surfaces of the flow path are coated with an alkylsilyl coating that is inert to at least one of the analytes. The alkylsilyl coating has the Formula I: R1, R2, R3, R4, R5, and R6 are each independently selected from (C1-C6)alkoxy, —NH(C1-C6)alkyl, —N((C1-C6)alkyl)2, OH, ORA, and halo. RA represents a point of attachment to the interior surfaces of the fluidic system. At least one of R1, R2, R3, R4, R5, and R6 is ORA. X is (C1-C20)alkyl, —O[(CH2)2O]1-20—, —(C1-C10)[NH(CO)NH(C1-C10)]1-20—, or —(C1-C10)[alkylphenyl(C1-C10)alkyl]1-20-.

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: 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.