English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Tailored methods for CGE-LIF based multiplexed high-throughput glycoanalysis with respect to sample characteristics

Borowiak, M., Hennig, R., Kottler, R., Reichl, U., & Rapp, E. (2011). Tailored methods for CGE-LIF based multiplexed high-throughput glycoanalysis with respect to sample characteristics. Glycoconjugate Journal, 28(5 ), 332.

Item is

Files

show Files
hide Files
:
347.pdf (Publisher version), 2MB
Name:
347.pdf
Description:
-
OA-Status:
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show

Creators

show
hide
 Creators:
Borowiak, M.1, Author           
Hennig, R.1, Author           
Kottler, R.1, Author           
Reichl, U.1, 2, Author           
Rapp, E.1, Author           
Affiliations:
1Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society, ou_1738140              
2Otto-von-Guericke-Universität Magdeburg, ou_1738156              

Content

show
hide
Free keywords: -
 Abstract: Glycomics is a rapidly emerging field, that can be viewed as a complement to other „omics“ approaches including proteomics and genomics. Hence, there is a dramatic increase in the demand for sophisticated databases and analytical tools in glycobiology, respectively glycobiotechnology. In order to enhance and improve the comparatively small set of existing glyco-analytical methods and toolboxes, a system and method for automated straightforward, sensitive high-throughput (HTP) and high-resolution glycoanalysis was developed [1,2]. This glycoanalysis approach is based on multiplexed capillary gelelectrophoresis with laser induced fluorescence detection (CGE-LIF), using a capillary DNA-Sequencer (Applied Biosystems). Data is evaluated in conjunction with a novel modular software-tool for data-processing and automated structural elucidation by interfacing a corresponding oligosaccharide-database. The aim of the project presented was to develop tailor-made workflows for multiplexed CGE-LIF based glycoanalysis, optimized for different types of samples, each with its specific characteristics. Exemplarily, three different sample types were chosen: first, a broad variety of N-glycans released from citrate plasma proteins (a complex sample of medium to large, charged and uncharged glyco-structures); second, a relative small set of N-glycans released from IgG (a modest sample with medium sized mainly uncharged glycostructures); and third, the complete pool of oligosaccharides from human breast milk (complex sample of small charged and uncharged glycostuctures). The whole workflow starting from sample clean-up until CGE-LIF analysis using a standard capillary DNA Sequencer was optimized individually to each of the three representative samples. For sample preparation, different chromatographic techniques and materials were tested and different polymers and capillary lengths were investigated for final CGE-LIF analysis.
[1] Schwarzer, J.; E. Rapp; U. Reichl; Electrophoresis (2008) 29, 4203-4214.
[2] Ruhaak, L.R.; Hennig, R.; Huhn, C.; Borowiak, M.; Dolhain, R. J. E. M.; Deelder, A. M.; Rapp, E.; Wuhrer, M.; Journal of Proteome Research (2010) 9, 6655–6664.

Details

show
hide
Language(s): eng - English
 Dates: 2011
 Publication Status: Issued
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 571752
DOI: 10.1007/s10719-011-9334-5
 Degree: -

Event

show
hide
Title: 21st International Symposium on Glycoconjugates - Glyco 21
Place of Event: Wien, Östereich
Start-/End Date: 2011-08-21 - 2011-08-26

Legal Case

show

Project information

show

Source 1

show
hide
Title: Glycoconjugate Journal
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 28 (5 ) Sequence Number: - Start / End Page: 332 Identifier: -