Abstract:

[Abstract] Objective To prepare cetuximab-β-glucosidase conjugates and to identify its enzymatic activity and an? tibody activity. Methods Cetuximab and β-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 of β-glucosidase, cetuximab, cetuximab-β-glucosidase conjugates through electropherogram. Although the en? zymatic activity of cetuximab-β-glucosidase conjugates was lower than that of β-glucosidase (U/L： 672.97±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.

Key words: Cetuximab, β-glucosidase, Conjugates, urinary bladder neoplasms, Sulfo-SMCC, carcinoma