Preparation and characterization of a single chain antibody fragment of mAb SZ-21 against platelets GPIIIa.
- Author:
Guangyu AN
1
;
Ningzheng DONG
;
Bojing SHAO
;
Mingqing ZHU
;
Changgeng RUAN
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; pharmacology; Blood Platelets; metabolism; Endothelium, Vascular; cytology; Fibrinogen; metabolism; Humans; Immunoglobulin Fragments; pharmacology; Platelet Aggregation; Platelet Glycoprotein GPIIb-IIIa Complex; immunology; Recombinant Fusion Proteins; isolation & purification; pharmacology
- From: Chinese Journal of Hematology 2002;23(9):480-482
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare a single chain antibody (ScFv) of mAb SZ-21 against platelet GPIIIa for its future clinical application.
METHODSThe expression vector pET20b-SZ-21ScFv was constructed and the fusion protein was expressed in E. coli BL21 (DE3) PlysS. The activated fusion protein was obtained after a series of purification steps, including cell breakage, inclusion body solubilization, His-bind resin affinity chromatography and protein refolding.
RESULTSThe fusion protein yields were up to 21% of the total amount of bacteria protein. The ScFv fragment could inhibit ADP-induced platelets aggregation in a dose-dependent manner in vitro and the maximal inhibition rate was obtained at a concentration of 20 micro g/ml. It also reacted with endothelial cells as detected by flow cytometry. Moreover, the ScFv fragment was able to inhibit the binding of fibrinogen to platelet.
CONCLUSIONThe SZ-21ScFv fragment had the activity to inhibit platelets aggregation and the binding of fibrinogen to platelet, being potentially useful for the treatment of thrombotic diseases.
