Green Fluorescent Protein-Based System for Analysis of E-Selectin-Mediated Adhesion. (March 1999)

• Record Type: Journal Article
• Authors: El-Battari, A.; Zerfaoui, M.; Panicot, L.; Mas, E.; Prévôt, C.; Lombardo, D.
• Is Part Of: Biotechniques. Volume 26:Number 3(1999)
• Page Start: 536
• Page End: 540
• Publication Date: 1999-03
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/99263cr03
• Languages: English
• ISSNs: 0736-6205

Abstract

Numerous cell-based or cell-free systems for study of selectin adhesion use radiolabeled tracers. However, in addition to handling problems associated with the use of radioisotopes, these assays have difficulty relating a number of counts to a number of adherent cells. Here, we describe an assay that uses the natural fluorescence of the green fluorescent protein (GFP) to measure binding of cells to E-selectin. We elaborated an adhesion system composed of a cell monolayer expressing E-selectin ligand to which monodispersed fluorescent Chinese hamster ovary (CHO) cells expressing E-selectin are added. Due to GFP autofluorescence, adhered cells can be easily distinguished from cell monolayers by fluorescence microscopy, and adhesion can be measured by cytofluorometry. We applied this GFP-based adhesion assay to measure the adherence of a pancreatic tumor cell line and found that the binding parameters of these cells satisfy a number of E-selectinspecific criteria.