Agilent Technologies, Inc.

Sierra Sensors Gmbh

Karla M Robotti

Daniel Bernardo Roitman

Hongfeng Yin

Sally A Swedberg

Kevin P Killeen

James Alexander Apffel, Jr

Karin Caldwell

Karen L Seaward

Magdalena Anna Ostrowski

Rudolf Grimm

Microfluidic devices and methods for analyzing glycan profiles of glycoproteins are provided. Some embodiments of the devices comprise a deglycosylation column for cleaving glycans, an optional cleaning column for removing proteins, a trapping column for enriching glycans, and a separation column for resolving glycans. The devices and methods significantly improve the speed and sensitivity of glycan analysis.

Microfluidic glycan analysis

A microfluidic system including a chip sensor and an SPR optical detector. The chip sensor may be made a non-transparent material, such as polyimide or silicon, allowing non-visible radiation produced by the SPR optical detector to pass through and interact with a surface plasmon generating layer on the chip sensor.

Microfluidic devices with SPR sensing capabilities

Aspects of the invention include a microfluidic device having a fluid flow path and a monolithic chromatographic material covalently bound to at least a portion of a polymeric surface of the fluid flow path. Aspects of the invention further include methods of making and using such devices.

Microfluidic devices comprising fluid flow paths having monolithic chromatographic material

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 invention in particular embodiments provides an evanescent wave sensor which includes a ligand bound to a sensor substrate via a NCYX linker moiety, as defined herein. Methods of making the subject evanescent wave sensors are also provided which include contacting a first reactive moiety which has an isocyanato (or isothiocyanato) moiety with a second reactive moiety which has an hydroxyl, thiol, or amino moiety, as further described herein. Also provided by the invention are methods in which a subject evanescent wave sensor is contacted with a sample, and binding of analytes in the sample to the sensor is assessed by evanescent wave detection. The invention also provides kits and systems for performing the subject methods.

Evanescent wave sensor with attached ligand

A method useful in the analysis of glycosylated proteins, in which a mixture containing glycosylated proteins and unglycosylated proteins is contacted with a resin that includes a nucleophile bound to a solid support via a linker. The contacting is performed under conditions sufficient to result in removal of the glycosyl group from the glycosylated proteins and to concomitantly result in the deglycosylated proteins covalently bound to the solid support. The deglycosylated proteins bound to the solid support may be rinsed to remove proteins that are not covalently bound to the solid support. The deglycosylated proteins are released from the solid support and may be subjected to further purification and/or analysis.

Enrichment and tagging of glycosylated proteins

A fabrication and adhesion method for a polyaryl-ether-ketone (PAEK) device, such as a microfluidic device, is disclosed. At least one glassy uncrystallized PAEK substrate is heated up to near or above the glass transition temperature to allow the substrate to crystallize from the glass state, while embossing the substrate with patterns. Bonding the PAEK substrate to another substrate is accomplished using a solvent-resistant adhesive, such as a polyimide-based adhesive, in combination with an adhesion enhancement treatment. In certain embodiments, the adhesion enhancement treatment is a plasma treatment or a chemical sulfonation treatment.

Paek embossing and adhesion for microfluidic devices

The present invention is a method, and a kit for carrying out the method, of separating phosphorylated peptides from a mixture of phosphorylated and unphosphorylated peptides. The method involves reacting a collection of peptides, in which some of the peptides have one or more phosphate groups, with a first resin. The peptides are then reacted with a capture ligand which reacts with the phosphate group of the phosphorylated peptides to form a bond. The capture ligand is used to separate the phosphorylated peptides from the unphosphorylated peptides.

Enrichment of phosphate peptides for proteomic analysis

Methods and compositions for modifying the surface of a polymeric substrate are provided. In the subject methods, at least a portion of the surface of the polymeric substrate is contacted with a biofouling resistant surfactant composition. At least a portion of the surfactant molecules of the composition are end modified with a non-reactive, charged end group that is charge stable over a pH range of about 2 to 12. The subject methods and compositions find particular use with electrophoretic devices, such as capillary electrophoretic devices.

Methods and compositions for modifying the surface of a polymeric substrate

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

Methods for selectively separating at least one component from a multi-component componant fluidic sample are provided. In the subject methods, the fluidic sample is introduced into a micro-fluidic device that includes at least one micro-valve made up of a phase reversible material. The multi-component fluidic sample is then contacted with the microvalve in a microfluidic device under conditions sufficient for the at least one component to enter the microvalve, while the remaining constituents of the fluidic sample remain outside of the microvalve. Also provided are kits for use in practicing the subject methods, where the kits include at least a microfluidic device having a microvalve and instructional material (or means for obtaining the same) on how to use the device in the subject methods. The subject devices find use in a variety of applications, including sample desalting and concentration applications.

Methods and using chemico-mechanical microvalve devices for the selective separation of components from multi-component fluid samples

Methods and compositions for modifying the surface of a polymeric substrate are provided. In the subject methods, at least a portion of the surface of the polymeric substrate is contacted with a biofouling resistant surfactant composition. At least a portion of the surfactant molecules of the composition are end modified with a non-reactive, charged end group that is charge stable over a pH range of a bout 2 to 12. The subject methods and compositions find particular use with electrophoretic devices, such as capillary electrophoretic devices.

Micro-fluid devices and methods for their use are provided. The subject devices are characterized by the presence of at least one micro-valve comprising a phase reversible material, e.g. a reversible gel, that reversibly changes its physical state in response to an applied stimulus, e.g. a thermoreversible gel. In using the subject device, fluid flow along a flow path of the device is modulated by applying an appropriate stimulus, e.g. changing the temperature, to the microvalve. The subject devices find use in a variety of applications, including micro-analytical applications.

Growing community of inventors

Cupertino, CA, United States of America

Karla M Robotti