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Binding and orientation of fibronectin to silanated glass surfaces using immobilized bacterial adhesin-related peptides

Paper ID Volume ID Publish Year Pages File Format Full-Text
12327 790 2003 8 PDF Available
Title
Binding and orientation of fibronectin to silanated glass surfaces using immobilized bacterial adhesin-related peptides
Abstract

Previously, we have demonstrated the suitability of bacterial adhesin-related peptides, directly immobilized on polystyrene surfaces, to bind and orient fibronectin (FN). For these studies a method to bind the large protein FN in a desired orientation on a solid substratum was developed which utilizes a bacterial adhesin-related peptide (designated BRP-A), which is known to bind specifically to the NH3-terminus end of FN. Glass substrata was first coated with an amine-terminated silane, followed by streptavidin (SA), which was used as an intermediate tether to bind the biotinylated bacterial adhesin-related peptide. The BRP-A peptide, used for these studies was synthesized with a terminal biotin to assure irreversible coupling of the BRP-A to the streptavidin. The biotinylated BRP-A was next immobilized on the SA-silanated glass surfaces. 125I-FN was used to quantify the amount of FN binding to the (BRP-A):SA-silanated glass surface. Monoclonal antibodies, which react with specific epitopes at either the NH3– or –COOH-termini of FN, were used to quantify the binding and orientation of FN. The Results of these studies indicated: (1) FN bound to the BRP-A:SA-silanated glass surface; and (2) the bound FN was oriented such that NH2-terminal region of FN was bound towards the glass surface and the COOH-terminus was oriented away from the glass surface. These studies demonstrate that small peptides can be used to specifically bind and orient large proteins such as FN on the surfaces.

Keywords
Fibronectin; Orientation; Bacterial-related peptides; Anti-fibronectin antibodies; Silanized surfaces
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Binding and orientation of fibronectin to silanated glass surfaces using immobilized bacterial adhesin-related peptides
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 24, Issue 22, October 2003, Pages 3877–3884
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us