Microscale exoglycosidase processing and lectin capture of glycans with phospholipid assisted capillary electrophoresis separations

Details

• Personal Author:
  Archer-Hartmann SA ; Holland LA ; Lowry DT ; Sargent LM
• Description:
  Capillary electrophoresis separations of glycans labeled with l-aminopyrene-3,6,8-trisulfonic acid were achieved with separation efficiencies ranging from 480 000 to 640 000 theoretical plates in a 60.2 cm, 25 mu m inner diameter fused silica capillary. Under these separation conditions, the coefficient of variation in peak area is 10%, and if labeling efficiency is estimated at 100%, the limit of detection is 15 fM. The capillary electrophoresis method incorporated phospholipid additives to enhance the separation of glycans with slight differences in hydrodynamic volume. In addition, the phospholipid additives supported the integration of the lectin concanavalin A as well as the enzymes alpha 1-2,3 mannosidase or beta 1-4 galactosidase to provide structural and compositional information about the glycans subject to separation. The use of in-capillary cleavage of terminal glycan residues with exoglycosidases offers a number of advantages over benchtop enzymatic sequencing, including reduced consumption of analyte, as well as enzyme. These methods were used to evaluate glycans derived from the glycoproteins alpha 1-acid glycoprotein, fetuin, and ribonuclease B, as well as from glycoproteins collected from MCF7 cells. [Description provided by NIOSH]
• Subjects:
  Chemistry Chemistry, Analytic Microbiology Occupational Health Safety
• Keywords:
  Analytical-chemistry Analytical-processes Biological-effects Chemical-analysis Chemical-composition Chemical-processing Exposure-levels Mathematical-models Microscopic-analysis Monitoring-systems Quantitative-analysis Statistical-analysis

  More +
• ISSN:
  0003-2700
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  83
• Issue:
  1
• NIOSHTIC Number:
  nn:20038612
• Citation:
  Anal Chem 2011 Apr; 83(7):2740-2747
• Contact Point Address:
  Lisa A Holland, C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506
• Email:
  lisa.holland@mail.wvu.edu
• Federal Fiscal Year:
  2011
• NORA Priority Area:
  Manufacturing
• Peer Reviewed:
  True
• Source Full Name:
  Analytical Chemistry
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:447b3e24a6c810d1aec698ce0618094c80a300f9d2271e6d8c17f92475197884380364964dbd0881305759417fb66cb3ca17dd3cc66d7e305a2e7869f1d53bf0
• Download URL:
  https://stacks.cdc.gov/view/cdc/192263/cdc_192263_DS1.pdf
• File Type:
  [PDF - 1.16 MB ]