Abstract/Details

A hydrodynamic microfluidic system for the analysis of glycoproteins


2006 2006

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Abstract (summary)

An integrated device has been described to perform a variety of enzymatic and immunological assays for applications to proteomics, disease diagnosis, and disease state monitoring. The work presented here focused on the use of this system to perform solution enzyme assays and lectin immunoassays.

In typical microfluidic devices, fluid manipulation is conducted via microfabricated pumps or electroosmotic flow. On such a small scale, pumps and valves are extremely difficult to fabricate and have a high failure rate. Electroosmotic flow requires expensive power supplies, incurs a sampling bias, and can result in joule heating within plastic devices. A plastic device was proposed that employed only hydrodynamic flow for fluid transport. This device was used to carry out enzymatic assays for two small molecule analytes. Several coating strategies were considered to prevent fouling of the device as a result of adsorption of both large and small molecules to the hydrophobic walls.

Post-translational modifications such as glycosylation and phosphorylation are of great interest in proteomics, and high-throughput methods are needed for the identification of trends in these modifications. An immunoassay was developed for the characterization of glycoproteins. Antibodies were used to capture target glycoproteins, and lectins used for qualitative and quantitative analysis of the glycans present on the protein, The assay was carried out in a traditional format in microtitre wells and in a microscale format with microcontact printing of assay arrays. The assay successfully distinguished between types of glycosylation on two standard proteins, and quantitation was achieved by two independent detection methods. It was also determined that the hydrodynamic flow system was an effective means of concentration of low-abundance proteins on the assay.

The lectin immunoassay was adapted to investigate two known protein markers for rheumatoid arthritis. A variety of lectins were used to characterize the types of glycosylation on both standard proteins, and a comparative study was performed to identify differences in glycosylation between samples of serum from a normal pool and a rheumatoid arthritis patient. This assay successfully identified trends in glycosylation that are consistent with those reported in the literature.

Indexing (details)


Subject
Analytical chemistry
Classification
0486: Analytical chemistry
Identifier / keyword
Pure sciences; Glycoproteins; Hydrodynamic; Lectin; Microfluidic; Proteomics
Title
A hydrodynamic microfluidic system for the analysis of glycoproteins
Author
Miller, Elizabeth Marie
Number of pages
220
Publication year
2006
Degree date
2006
School code
0183
Source
DAI-B 67/09, Dissertation Abstracts International
Place of publication
Ann Arbor
Country of publication
United States
ISBN
9780542864957
Advisor
Regnier, Fred E.
University/institution
Purdue University
University location
United States -- Indiana
Degree
Ph.D.
Source type
Dissertations & Theses
Language
English
Document type
Dissertation/Thesis
Dissertation/thesis number
3232214
ProQuest document ID
305265094
Copyright
Database copyright ProQuest LLC; ProQuest does not claim copyright in the individual underlying works.
Document URL
http://search.proquest.com/docview/305265094/abstract
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