Preparation and Identification of Cetuximab-β-Glucosidase Conjugates
10.3969/j.issn.0253-9896.2014.12.001
- VernacularTitle:西妥昔单抗-β葡萄糖苷酶偶联物的制备及鉴定
- Author:
Yunlong LIU
;
Jie ZHOU
;
Guangwen WANG
;
Zhen NIE
- Publication Type:Journal Article
- Keywords:
cetuximab;
β-glucosidase;
conjugates;
urinary bladder neoplasms;
carcinoma;
Sulfo-SMCC
- From:
Tianjin Medical Journal
2014;(12):1153-1155
- CountryChina
- Language:Chinese
-
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/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.
