Preparation and Identification of Cetuximab-β-Glucosidase Conjugates / 天津医药

ABSTRACT

Objective To prepare cetuximab-β-glucosidase conjugates and to identify its enzymatic activity and an?tibody activity. Methods Cetuximab andβ-glucosidase were crosslinked by Sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC). Cetuximab-β-glucosidase conjugates and its enzymatic and antibody activity were examined by non-reduced SDS-PAGE, colorimetry and indirect immunofluorescence assay. Results We can see clear bands ofβ-glucosidase, cetuximab, cetuximab-β-glucosidase conjugates through electropherogram. Although the en?zymatic activity of cetuximab-β-glucosidase conjugates was lower than that ofβ-glucosidase (U/L672.97±46.19 vs 869.50± 57.28,t=5.972,P<0.05) shown by colorimetry assay, it still maintain good enzymatic activity. Under fluorescence micro?scope, we can see the conjugates interacted with human bladder cancer EJ cells are in a red fluorescence. Conclusion Ce?tuximab,β-glucosidase were crosslinked successfully by Sulfo-SMCC without altered its enzymatic and antibody activity.