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Glycans

Glycans

Research in the Lebrilla Clinical Glycomic Laboratory has focused on methods for the determination of oligosaccharide structures in various glyconjugates including free oligosaccharides, glycolipids and glycoproteins. Analysis of glycans in this laboratory is performed on several levels commensurate with the complexity of glycosylation.   Rapid throughput structural identification is performed with milk oligosaccharides and N-glycans, particularly those from serum.  Site-specific glycosylation is determined for glycoproteins in mixtures or pure.  Glycolipids profiling is performed on gangliosides.

 

The lab has developed a glycan structural database with hundreds of structures that characterized by accurate mass, LC retention time, and tandem MS. Typically, human and animal body fluids are analyzed.  The lab currently supports researchers innutri-glycomics in collaboration with the Milk Genomics Consortium and in biomarker discovery for cancer and infectious disease through the characterization of dysregulatedglycans.

 

glycans1

 

 

 

 

glycans2

 

 

 

 

 

 

 

 

 

 

Sample Glycan Report

We will build further these successes with the following aims for the Laboratory:

 

  1. To increase throughput of quantitative glycan profiling of free oligosaccharides in milk, urine, feces and blood by developing a rapid identification method employing retention times, accurate mass and tandem MS. We aim to accurately profile 50 samples per day while tracking as many as 300 structures simultaneously.
  2. To increase throughput of N-glycan profiling in serum and plasma using the same approach for milk oligosaccharides based on a library of N-glycan structures.
  3. To develop and employ a target MRM-library for LC-triple quadrupole mass spectrometry after discovery works in LC-QTOF MS for accurate glycan quantitation.
  4. To perform glycan analysis using dried blood spot technology. Fluids on filter paper are excellent options for obtaining samples from areas of the world without proper refrigeration, as well for newborn screening.
  5. To improve throughput for protein-specific glycosylation with site-specific glycan heterogeneity. This capability will provide, for the first time, unbiased examination of alterations in glycosylation on a protein and site-specific level.

 

The development of such assays will take considerable time, but would be an excellent result for the Center and a great service opportunity when the Center is fully self-sustained.

 

Equipment

Ion Spec MALDI-FTICR mass spectrometer from Ionspec Inc. with a 7 Tesla superconducting magnet.

Varian ESI Triple Quad FTICR mass spectrometer with a 9.4 Tesla superconducting magnet.

Agilent 6490 Triple Quadrupole with ion funnel and Agilent UHPLC 1260.

Agilent 500 Ion Trap with Capillar LC.

Agilent 6500Chip-TOF with nanoliquid chromatography.

Agilent 6540 Chip-QTOF with nanoliquid chromatography.

Agilent 1100 HPLC with diode array detector.

Agilent 1200 capillary electrophoresis with diode array detector.

Gilson GXS 274 Aspec robotic liquid handler fraction collector.

 

 

Staff

Dr. Carlito B. Lebrilla (PI):cblebrilla@ucdavis.edu • Phone: 530-752-6364 • Fax: 530-752-8995

Dr. L. Renee Ruhaak (Postdoc):  lruhaak@ucdavis.edu• Phone: 530-752-5504 • Fax: 530-752-8995

Ms. Lauren Dimpasoc (Lab Tech)lmdimapasoc@ucdavis.edu• Phone: 530-752-5504 • Fax: 530-752-8995

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