Related Experiment Videos
Human single chain antibodies against heparin: selection, characterization, and effect on coagulation
Els M A van de Westerlo1, Toon F C M Smetsers, Michel A B A Dennissen
1Department of Biochemistry, University Medical Center Nijmegen, 6500 HB Nijmegen, The Netherlands.
Blood
|March 16, 2002
Summary
Researchers developed 19 human anti-heparin antibodies using phage display. These antibodies are valuable tools for studying heparin and related glycosaminoglycans (GAGs), with some inhibiting heparin
Area of Science:
- Biochemistry
- Immunology
- Pharmacology
Background:
- Heparin, a glycosaminoglycan (GAG), is crucial as an anticoagulant and is found in mast cells and basophilic granulocytes.
- Understanding heparin's structure and function is vital for its therapeutic applications and for developing related treatments.
Purpose of the Study:
- To select and characterize unique human antibodies against heparin.
- To investigate the specificity and epitope mapping of these anti-heparin antibodies.
- To evaluate the potential of these antibodies as tools for studying GAGs and as therapeutic agents.
Main Methods:
- Phage display technology was employed to select 19 unique human anti-heparin antibodies.
- Antibody specificity was assessed against heparin, heparan sulfate, and chondroitin sulfate, with emphasis on the role of sulfate groups.
- Immunohistochemical staining on tissue cryosections and enzymatic digestion were used to determine antibody localization and GAG specificity.
- The impact of antibodies on heparin's anticoagulant activity was evaluated using the activated partial thromboplastin time (APTT) assay.
Main Results:
- 19 distinct human anti-heparin antibodies were successfully selected.
- Antibodies exhibited varying specificities for different GAGs, with sulfate groups being essential for binding.
- Antibodies demonstrated specific staining patterns in rat tissues, confirming their reactivity with heparin in situ.
- Three antibodies significantly inhibited heparin's anticoagulant activity in vitro, with efficacy linked to heparin binding.
- Some antibodies recognized the heparin pentasaccharide crucial for antithrombin III interaction.
Conclusions:
- The selected human antibodies are specific tools for investigating the structure, localization, and function of heparin and related GAGs.
- The antibodies' ability to inhibit heparin's anticoagulant activity suggests potential applications as blocking agents.
- Further research utilizing these antibodies can advance our understanding of GAG biology and anticoagulant mechanisms.