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Preparation of Anti-CEA McAb-β-Glucosidase Conjugate and Its Killing Effect Against LoVo Cells in Combination with Amygdalin in vitro
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Abstract:
Objective: To prepare anti-CEA monoclonal antibody-β-glucosidase conjugate(Anti-CEA McAb-β-glucosidase conjugate)， evaluate its immunoreactivity and enzymatic activity, and study its killing effect against LoVo cells in combination with amygdalin in vitro.Methods: Anti-CEA monoclonal antibody-β-glucosidase conjugate was prepared to use heterobifunctional crosslinking reagent 3-maleimidobenzoic acid N-succinimidyl ester (MBS) conjugating β-glucosidase to a anti-CEA monoclonal antibody, the immunoreactivity and enzymatic activity of the conjugate was tested by ELISA method. The cytotoxicity to targeted cells and untargeted cells following specific activation of amygdalin by the conjugate was evaluated by MTT assy in vitro.Results: Anti-CEA McAb-β-glucosidase conjugate kept both immunoreactivity and enzymatic activty. Anti-CEA McAb-β-glucosidase conjugate could bird LoVo cells specifically and activate amygdalin to kill Love cells in a dose-dependent manner, but it had not the same effect against MCF-7 cells.Conclusion: Anti-CEA McAb-β-glucosidase conjugate has a good character for binding LoVo cells in vitro and can activate amygdalin prodrug to kill targeted cells specifically.This way might become a hopeful model of colorectal carcinoma immuonotherapy.
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