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Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Published on: May 4, 2012
Developmental changes in heparan sulfate expression: in situ detection with mAbs.
G David1, X M Bai, B Van der Schueren
1Center for Human Genetics, University of Leuven, Belgium.
The Journal of Cell Biology
|November 1, 1992
Summary
Two novel monoclonal antibodies (mAbs) reveal the developmental distribution of heparan sulfate (HS) in tissues. These antibodies, 10E4 and 3G10, highlight HS presence during morphogenesis and indicate its developmentally regulated expression.
Area of Science:
- Biochemistry
- Developmental Biology
- Glycobiology
Background:
- Heparan sulfate (HS) is a crucial glycosaminoglycan involved in various biological processes.
- Understanding the spatiotemporal distribution of HS is essential for comprehending its developmental roles.
- Existing methods for HS analysis have limitations in distinguishing specific epitopes.
Purpose of the Study:
- To generate and characterize novel monoclonal antibodies (mAbs) for analyzing HS distribution.
- To investigate the cellular and tissular localization of HS during development using these mAbs.
- To explore the structural basis and developmental regulation of HS expression.
Main Methods:
- Development of two specific mAbs, 10E4 and 3G10, targeting distinct HS epitopes.
- Enzymatic treatments (heparitinase, heparinase) to characterize antibody epitope specificity.
- Immunohistochemical staining of embryonic and adult tissues to determine HS distribution.
- Analysis of antibody reactivity with native and modified HS chains and proteoglycans.
Main Results:
- mAb 10E4 recognizes native HS, while mAb 3G10 recognizes a heparitinase-generated, desaturated uronate epitope.
- Both antibodies show HS localization on cell surfaces, extracellular matrix, and basement membranes.
- Significant temporal and regional variations in HS epitope abundance were observed during development.
- Differential staining patterns suggest regional differences in HS structure or exposure.
Conclusions:
- Heparan sulfate is abundant at sites of active morphogenesis.
- The expression and presentation of HS are developmentally regulated.
- These novel mAbs provide valuable tools for studying HS in developmental contexts.
