Evaluation of allergen during processes of Fufang Kushen injection by IgG-promoter-HepG2 cell assay in vitro.
- Author:
Zhiyi YUAN
1
;
Yugang WANG
;
Fan LEI
;
Xi LU
;
Shuang ZHAO
;
Jingfei JIANG
;
Huiyu LI
;
Bohang CHEN
;
Zhimin WANG
;
Dongming XING
;
Yi DING
;
Lijun DU
Author Information
- Publication Type:Journal Article
- MeSH: Allergens; analysis; immunology; Drugs, Chinese Herbal; administration & dosage; chemistry; Gene Expression Regulation; immunology; Hep G2 Cells; Humans; Immunoglobulin G; genetics; Injections; Medicine, Chinese Traditional; standards; Promoter Regions, Genetic; genetics; Quality Control
- From: China Journal of Chinese Materia Medica 2011;36(14):1850-1854
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro.
METHODBy transfecting a IgG promoter-regulating the expression of green fluorescent protein(GFP) plasmid into HepG2 cells, this transferred cells were incubated with common allergens (like puerarin, ovalbumin, LPS or Sal typhoid vi polysaccharide vaccine), excipients using in Fufang Kushen injection (NaOH, acetic acid, Tween-80 and ethanol) and samples from the key production processes of the injection for 30 minutes . Fluorescent photographs were analyzed the fluorescence intensity of the cells by using an image analysis software.
RESULTAll of common allergens significantly increased the IgG expression. Two of the excipicents, acetic acids and Tween-80 were shown to increased the IgG expression, while others had no effect on IgG expression. In the 8 samples from the key processes in the production of Fufang Kushen injection, two of them stimulated IgG expression.
CONCLUSIONIgG promoter-HepG2 cells are highly sensitive and specific to allergens, and thus can be applied to rapid screening of allergens in components and injections in transcriptional level. It is possible to use the IgG-promoter HepG2 cells in a real-time monitoring of allergens in the production processes of Chinese medicine injections.