Preparation and Activity of Monoclonal Antibody against Intercellular Adhesion Molecule-1

Objective To prepare the monoclonal antibody(McAb) against human intercellular adhesion molecule-1(ICAM-1) with biological activity.Methods Immunize BALB/c mice with human ICAM-1,then screen the hybridoma cell strain stably secreting McAb against ICAM-1 by selective culture with HAT and clone by limiting dilution.The secreted McAb was purified by caprylic acid-ammonium sulfate precipitation and affinity chromatography,then determined for subclass and titer by indirect ELISA,for specificity by Western blot and for neutralizing activity by cell adhesion test.Results A hybridoma cell strain 3F2 stably secreting McAb against ICAM-1,with a chromosome modal number of 98～104,was screened.The secreted McAb was IgG1 and reached a protein content of 1.253 mg/ml,a purity of 83.6% and a titer of 1:2.56×105 after purification.The purified McAb bound to ICAM-1 specifically,inhibited the adhesion of ICAM-1 to leucoma cells and showed significant neutralizing activity to ICAM-1.Conclusion The McAb against ICAM-1 was successfully prepared,which laid a foundation of further study on its application.