Journal: The Journal of Biological Chemistry
Article Title: Identification, Characterization, and Epitope Mapping of Human Monoclonal Antibody J19 That Specifically Recognizes Activated Integrin ? 4 ? 7 *
doi: 10.1074/jbc.M112.341263
Figure Lengend Snippet: The binding specificities of J19 phage and J19 scFv to integrin α4β7. A, monoclonal phage ELISAs were carried out with coated soluble WT α4β7, wedge mutant α4β7, and BSA control in the presence of indicated divalent cations. ELISA signals of J19 phage are shown. The error bars are ± S.D. (n = 3). B, 5 μg/ml J19 scFv or Act-1 binding to inactive (1 mm Ca2+ + 1 mm Mg2+) or active (2 mm Mn2+) α4β7 stably expressed on K562 cells were analyzed by flow cytometry. A representative experiment (of three) is shown as a histogram. The numbers within the panels show the specific mean fluorescence intensity of Act-1 and J19 scFv. The results are the means ± S.D. of three independent experiments. Open histogram, control mouse IgG or control scFv; filled histogram, Act-1 or J19 scFv.
Article Snippet: Selection of Integrin-binding Phages Human scFv phage library was purchased from Geneservice.
Techniques: Binding Assay, Mutagenesis, Enzyme-linked Immunosorbent Assay, Stable Transfection, Flow Cytometry, Fluorescence