Bi-ligand surfaces with oriented and patterned protein for real-time tracking of cell migration.

Published

Journal Article

A bioactive platform for the quantitative observation of cell migration is presented by (1) presenting migration factors in a well-defined manner on 2-D substrates, and (2) enabling continuous cell tracking. Well-defined substrate presentation is achieved by correctly orienting immobilized proteins (chemokines and cell adhesion molecules), such that the active site is accessible to cell surface receptors. A thiol-terminated self-assembled monolayer on a silica slide was used as a base substrate for subsequent chemistry. The thiol-terminated surface was converted to an immobilized metal ion surface using a maleimido-nitrilotriacetic acid (NTA) cross-linker that bound Histidine-tagged recombinant proteins on the surface with uniform distribution and specific orientation. This platform was used to study the influence of surface-immobilized chemokine SDF-1α and cell adhesion molecule ICAM-1 on murine splenic B lymphocyte migration. While soluble SDF-1α induced trans-migration in a Boyden Chamber type chemotaxis assay, immobilized SDF-1α alone did not elicit significant surface-migration on our test-platform surface. Surface-immobilized cell adhesion protein, ICAM-1, in conjunction with activation enabled migration of this cell type on our surface. Controlled exposure to UV light was used to produce stable linear gradients of His-tagged recombinant SDF-1α co-immobilized with ICAM-1 following our surface chemistry approach. XPS and antibody staining showed defined gradients of outwardly oriented SDF-1α active sites. This test platform can be especially valuable for investigators interested in studying the influence of surface-immobilized factors on cell behavior and may also be used as a cell migration enabling platform for testing the effects of various diffusible agents.

Full Text

Duke Authors

Cited Authors

  • Vernekar, VN; Wallace, CS; Wu, M; Chao, JT; O'Connor, SK; Raleigh, A; Liu, X; Haugh, JM; Reichert, WM

Published Date

  • November 2014

Published In

Volume / Issue

  • 123 /

Start / End Page

  • 225 - 235

PubMed ID

  • 25262410

Pubmed Central ID

  • 25262410

Electronic International Standard Serial Number (EISSN)

  • 1873-4367

International Standard Serial Number (ISSN)

  • 0927-7765

Digital Object Identifier (DOI)

  • 10.1016/j.colsurfb.2014.09.020

Language

  • eng