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A 2D Covalently Bound Continuous Protein Gradient Assay

Published online by Cambridge University Press:  22 January 2014

Benjamin Mintz and
James A. Cooper Jr.
Benjamin Mintz
Affiliation:
Musculoskeletal &Translational Tissue Engineering Research (MATTER) Lab, Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th St, Troy, NY 12180, U.S.A.
James A. Cooper Jr.
Affiliation:
Musculoskeletal &Translational Tissue Engineering Research (MATTER) Lab, Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 110 8th St, Troy, NY 12180, U.S.A.
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Abstract

Traditional methods of quantifying cell movement in response to a chemotactic factors provide either a binary count of cell migration in response to a known concentration of the factor of interest in solution, as in Boyden chamber assays, or a method of tracking cells to determine velocities across a solubilized protein gradient where exact concentrations vary over time and are difficult to define, as in the Ibidi chemotaxis gradient assay. Using a silane self-assembling monolayer (SAM)-based procedure pioneered by V Hlady and associates, we have developed an assay capable of covalently binding a wide variety of proteins to an optically transparent surface in a 2D pattern via amine linkages. The pattern was then verified by contact angle and Raman and X-ray photoelectron spectroscopy. This new assay provides greater control of protein concentration and gradient intensity than when using only solubilized proteins.

Keywords

biomedicalsurface chemistryself-assembly

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Type
Articles
Information
MRS Online Proceedings Library (OPL) , Volume 1623: Symposium F – Synthetic Tools for Understanding Biological Phenomena , 2014, mrsf13-1623-f07-04-i08-04
Copyright
Copyright © Materials Research Society 2014 

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