Agilent Technologies, Inc.

James Alexander Apffel, Jr

James L Bertsch

Paul C Goodley

Mark H Werlich

Karla M Robotti

Brian Phillip Smart

Kent D Henry

Steven Michael Fischer

Hongfeng Yin

Arthur Schleifer

Mel N Kronick

Kristin Briana Bernick

Thomas P Doherty

Karen M Hahnenberger

Christine A Miller

Genevieve Van De Bittner

Brooks Bond-Watts

Martin Verhoef

Reagents and methods for extracting and stabilizing metabolites at room temperature and methods in which metabolites and one or more of proteins, lipids and nucleic acids are extracted from a single sample in a unified workflow.

Room temperature methods for preparing biological analytes

Provided herein is a method comprising one or more of the following steps: (a) lysing cells of a biological sample and contacting the biological sample with an amount of ionic liquid sufficient to denature intracellular metabolic enzymes in the biological sample to produce a contacted cellular sample; (b) mixing the contacted cellular sample with an organic solvent to produce an ionic liquid-organic solvent composition; (c) mixing the contacted cellular sample with the organic solvent to produce a dispersed microdroplet ionic liquid-organic solvent composition; (d) contacting the ionic liquid-organic solvent composition with an ion exchange composition to produce a second ionic liquid-organic solvent composition; (d) separating the ionic liquid from the organic solvent; and (e) extracting metabolites from the ionic liquid. Kits and systems for practicing the subject methods are also provided.

Method for metabolomic sample preparation based on ionic liquid dispersive liquid-liquid microextraction

A method for removing an ionic liquid from an aqueous sample is provided. In some embodiments, the method includes: (a) combining an aqueous sample including an ionic liquid with an ion exchanger composition including an ion exchanger counterion to produce a solution including a fluorous salt of the ionic liquid, where at least one of the ionic liquid and the ion exchanger counterion is fluorinated; (b) contacting the solution with a fluorous affinity material, thereby removing fluorous salt from the solution and producing an aqueous eluate; and (c) collecting the aqueous eluate. In certain embodiments, the method further includes: contacting a cell with an ionic liquid composition to lyse the cell and produce an aqueous sample; and contacting the aqueous sample with a reverse phase substrate, thereby adsorbing proteins and/or lipids of the cell on the substrate. Compositions, kits and systems for practicing the subject methods are also provided.

Fluorous affinity extraction for ionic liquid-based sample preparation

A sample droplet generator transforms a segmented array of sample material into a continuous stream of droplets containing analytes. The droplets may serve as a sample source for a wide range of detectors and analytical instruments. As one example, the droplets may be introduced into an ion source of a spectrometer that measures ions produced from the droplets or photons emitted from the droplets.

Sample droplet generation from segmented fluid flow and related devices and methods

Crosslinking reagents and methods for using the same for analysis of protein-protein interactions, are provided. The crosslinking reagents include a trifunctional scaffold that links two protein linking groups to each other and branches to link an affinity tag, where the protein linking groups can be fragmented from the scaffold. The distance between the two protein linking groups can be selected to crosslink two proteins of a protein complex via accessible amino acid residues. Also provided are crosslinked polypeptide compounds and kits that include crosslinking reagents. These reagents and methods find use in a variety of applications in which crosslinking of proteins in desired.

Crosslinking reagent for analysis of protein-protein interaction

Compounds, compositions, methods for sequencing proteins and peptides, and methods for identifying proteins and peptides in a mixture, are disclosed. Compounds of formula A-B-C wherein A is a nucleophilic reactive group, B is a detectable moiety capable of being isotopically labeled, and C is a charge replacement group, are used to label the peptides at the N-terminus or the C-terminus. The tagged peptides can then be analyzed by mass spectroscopy.

Triazine derivatives as universal peptide isotope tag reagents (U-PIT)

The present invention provides methods for analyzing a peptide or peptides of interest in a protein sample using a combination of a relatively generic isotope tag with a decoupled selection process, allowing simplified customization of the application with a single reagent. These methods comprise providing a first and a second protein sample; labeling the first protein sample with a first Universal Peptide Isotope Tag (U-PIT) reagent and the second protein sample with a second U-PIT reagent; separating the peptide of interest from the combined first and second protein samples; and determining the relative amount of the first U-PIT reagent and the second U-PIT reagent bound to the peptide or peptides of interest. The U-PIT label of the present inventive methods has the following general formulaA-B-Cwherein A is a nucleophilic reactive group, B is a detectable moiety that can be isotopically labeled, and C is a charge replacement group.

Modular isotope coding approach to proteomic analysis

A method and apparatus are disclosed wherein a plurality of electric fields and of orthogonal spray configurations of vaporized analyte are so combined as to enhance the efficiency of analyte detection and mass analysis. The invention provides reduced noise and increased signal sensitivity in both API electrospray and APCI operating modes.

Orthogonal ion sampling for APCI mass spectrometry

A transfer capillary and interface are provided that allow samples from multiple fluid streams to be introduced into a single mass analyzer. The transfer capillary has two or more channels and is placed between a first chamber, in which ions from the multiple fluid streams are generated, and a second chamber that is in fluid communication with a mass analyzer. The interface includes ionizers that generate ionized sample from each fluid stream and that direct the ionized sample toward the channels of the transfer capillary. The transfer capillary includes a multiplex selector that allows ions to flow through a selected subset of channels and sequentially changes which of the channels are included in the subset of channels through which ions flow.

Multiplexing capillary array for atmospheric pressure ionization-mass spectrometry

The invention provides a microanalytical device for analyzing a fluid sample containing at least one analyte molecule. The device is constructed from a substrate and a cover plate each having a substantially planar surface and a microchannel formed therein. The cover plate is placed over the substrate such that the cover plate microchannel is arranged in opposing relationship with the substrate microchannel. A membrane is interposed between the substrate and the cover and has at least one pore sized to allow passage of the analyte molecule from the substrate microchannel to the cover plate microchannel. An analyte altering moiety is attached to an interior surface of the pore and is capable of chemically altering the analyte molecule. Also disclosed are a method for chemically altering and transporting an analyte molecule in a fluid and a method for identifying a plurality of biomolecules.

Microanalytical device containing a membrane for molecular identification

A method is provided for the separation of nucleic acid samples. The method includes: providing a test mixture of a plurality of nucleic acid samples, wherein each sample is labeled with a spectrally detectable tag; applying the test mixture to a reversed phase solid support; eluting the mixture under partially denaturing conditions; and detecting spectrally resolved signals produced by the nucleic acid samples labeled with the detectable tags.

Growing community of inventors

Mountain View, CA, United States of America

James Alexander Apffel, Jr