Preparation and identification of monoclonal antibody against glycated hemoglobin
- VernacularTitle:糖化血红蛋白的单克隆抗体制备及鉴定
- Author:
Yuehui XIANG
1
;
Lingling LU
;
Feipei TU
;
Jiansheng CHEN
;
Xinwei ZHANG
Author Information
1. 浙江省瑞安市中医院眼科
- Keywords:
Glycosylated hemoglobin;
Monoclonal antibody;
Diabetic macular edema
- From:
China Modern Doctor
2015;(11):22-24
- CountryChina
- Language:Chinese
-
Abstract:
Objective Using hybridoma technique and screened hybridoma cell strains stably, efficiently secreted anti glycosylated hemoglobin monoclonal antibody to provide specific material for the development of glycosylated hemoglobin ELISA kit. Methods The immune antigen was prepared by maleimide method, multi-level immune mice by BALB/c,through cell culture fusion, screening of hybridoma cell culture medium HAT, ammonium sulfate salting out method and G protein chromatography. Monoclonal antibody subclasses were identified by monoclonal antibody subtype identi-fication Kit operation. Results Through cell fusion, screening and cloning culture, etc., the final selection screened 1 strain stably secreting specific antibody hybridoma cell line, named N5B4; cell culture supernatant liquid was 1:5000, ascites titer was 1:100 million; a standard curve to calculate the concentration in the sample human glycat ed hemoglobin was 99.2%. Conclusion After KET monoclonal antibody cell line to obtain a high specificity and high sen-sitivity of screening.
