OCMG Core Laboratory Services (October 2006)

The OCMG Core Laboratory is located on the OUHSC campus in Oklahoma City, in BRC-I, Room 413.

Telephone: 405-271-2765.

Services, training and consulting are performed by OCMG lab personnel on a fee and collaborative basis to be arranged on a per project basis. Fees are charged to partially recover costs of expendable supplies, instrument maintenance, and personnel salary. For information, please contact Dr. Christa Feasley (271-2765) or Dr. Chris West (271-2227, x1247).

1. Consulting

2. Training for use of instrumentation

3. Analytical Services

a) Monosaccharide composition analysis by Dionex HPAEC

Acid hydrolysis or methanolysis

HPLC

Quantitation

b) GAG disaccharide composition analysis using absorbance or fluorescence detection

Heparan sulfate- RP ion pairing HPLC

Chondroitin sulfate- HPAEC

c) Low-medium throughput screening of glycan and glycoconjugate profiles and differences

Glycan release (chemical or enzymatic) and reducing end fluorophore derivitization

Chromatography (amine absorption, Dionex, charcoal, ion exchange)

Traditional and isotope-coded MS-technologies (MALDI and on-line electrospray)

d) Proteomic analyses with focus on posttranslational modification

Protein ID

Peptide analysis with MALDI MS-MS characterization of fractions

General glycan structural studies

e) Glycan structural studies

Release and labeling, permethylation

MS and MS-MS based strategies

exoglycosidase sequencing

radiolabel studies

f) Microscale Lectin-affinity enrichments of glycoproteins, glycopeptides and glycans using semi-automated HPLC methods

3. Instrumentation available

a) Dionex DX-600 HPAEC system equipped with an autosampler, ED50 electrochemical detector, AD25 ultraviolet detector, flow desalter, and a Perkin-Elmer 610TR flow radiometric detector, and appropriate columns and standards, for analysis of monosaccharides and oligosaccharides. Fractions can be collected for further analysis.

b) Beckman System Gold HPLC systems (3) equipped with UV and fluorimetric detectors, for separating and detecting oligosaccharides and glycopeptides based on other chemistries. Fractions can be collected for further analysis.

c) Agilent 1100 LC/MSD System with an autosampler, diode array detector, and Bruker ion-trap mass spectrometer, for on-line MS detection and analysis of sugars.

d) Akta FPLC (Amersham Pharmacia Biotech/GE Healthcare) FPLC system with absorbance monitor and fraction collector system.

e) Bruker Ultraflex-2 MALDI-TOF-TOF for MS-MS studies to identify and characterize glycans and glycopeptides.

f) Hewlett-Packard 5890 Series II gas chromatograph with FID detector for monosaccharide identification and

methylation linkage analysis.

4. Personnel

Christa Feasley, Ph.D., postdoctoral associate

Wendy Ives, Technician

Chris West, Ph.D., Director