Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-29T07:45:24.125Z Has data issue: false hasContentIssue false
  • English
  • Français

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.
Get access

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
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 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Amyot, F., et al. ., A new experimental method for the evaluation of the release profiles of drug-loaded microbeads designed for embolisation. ITBM-RBM, 2002. 23(5): p. 285–289.CrossRefGoogle Scholar
Westrin, B.A., Axelsson, A., and Zacchi, G., Diffusion measurement in gels. Journal of Controlled Release, 1994. 30(3): p. 189–199.CrossRefGoogle Scholar
Reis, C.P., et al. ., Review and current status of emulsion/dispersion technology using an internal gelation process for the design of alginate particles. Journal of microencapsulation, 2006. 23(3): p. 245–257.CrossRefGoogle ScholarPubMed
Mehrotra, S., et al. ., Time Controlled Protein Release from Layer‐by‐Layer Assembled Multilayer Functionalized Agarose Hydrogels. Advanced functional materials, 2010. 20(2): p. 247–258.CrossRefGoogle ScholarPubMed
Bhatia, S.K., et al. ., Fabrication of surfaces resistant to protein adsorption and application to two-dimensional protein patterning. Analytical biochemistry, 1993. 208(1): p. 197–205.CrossRefGoogle ScholarPubMed
Liu, J.-F., et al. ., Fabrication of colloidal gold micro-patterns using photolithographed self-assembled monolayers as templates. Thin Solid Films, 1998. 327: p. 176–179.CrossRefGoogle Scholar
Webb, K., Hlady, V., and Tresco, P.A., Relative importance of surface wettability and charged functional groups on NIH 3T3 fibroblast attachment, spreading, and cytoskeletal organization. Journal of biomedical materials research, 1998. 41(3): p. 422.3.0.CO;2-K>CrossRefGoogle ScholarPubMed
Corum, L.E. and Hlady, V., Screening platelet–surface interactions using negative surface charge gradients. Biomaterials, 2010. 31(12): p. 3148–3155.CrossRefGoogle ScholarPubMed
Bhatia, S.K., Hickman, J.J., and Ligler, F.S., New approach to producing patterned biomolecular assemblies. Journal of the American Chemical Society, 1992. 114(11): p. 4432–4433.CrossRefGoogle Scholar
Dulcey, C.S., et al. ., Deep UV photochemistry of chemisorbed monolayers: patterned coplanar molecular assemblies. Science, 1991. 252(5005): p. 551–554.CrossRefGoogle ScholarPubMed
Mooney, J., et al. ., Patterning of functional antibodies and other proteins by photolithography of silane monolayers. Proceedings of the National Academy of Sciences, 1996. 93(22): p. 12287–12291.CrossRefGoogle ScholarPubMed
Blawas, A. and Reichert, W., Protein patterning. Biomaterials, 1998. 19(7-9): p. 595–609.CrossRefGoogle ScholarPubMed
Sugimura, H., et al. ., Surface potential microscopy for chemistry of organic self-assembled monolayers in small domains. Nanotechnology, 2004. 15(2): p. S69.CrossRefGoogle Scholar
Bhatia, S.K., et al. ., Immobilization of acetylcholinesterase on solid surfaces: chemistry and activity studies. Sensors and Actuators B: Chemical, 1991. 3(4): p. 311–317.CrossRefGoogle Scholar
Bhatia, S.K., et al. ., Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces. Analytical biochemistry, 1989. 178(2): p. 408–413.CrossRefGoogle ScholarPubMed
Routh, V.H. and Helke, C.J., A novel technique for producing antibody-coated microprobes using a thiol-terminal silane and a heterobifunctional crosslinker. Journal of neuroscience methods, 1997. 71(2): p. 163–168.CrossRefGoogle Scholar
Shriver-Lake, L.C., et al. ., Antibody immobilization using heterobifunctional crosslinkers. Biosensors and Bioelectronics, 1997. 12(11): p. 1101–1106.CrossRefGoogle ScholarPubMed
Xing, W.-L., et al. ., Portable fiber-optic immunosensor for detection of methsulfuron methyl. Talanta, 2000. 52(5): p. 879–883.CrossRefGoogle ScholarPubMed
Laboratories, S.R., Standard Raman spectra. 1976: Sadtler Research Laboratories.Google Scholar