[Evaluation of allergen during processes of Fufang Kushen injection by IgG-promoter-HepG2 cell assay in vitro].
OBJECTIVE
To study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro.


METHODS
By 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.


RESULTS
All 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.


CONCLUSIONS
IgG 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.
Protein Science Laboratory of Ministry of Education, Laboratory of Pharmaceutical Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China.
Zhiyi
Z
Yuan
Yugang
Y
Wang
Fan
F
Lei
Xi
X
Lu
Shuang
S
Zhao
Jingfei
J
Jiang
Huiyu
H
Li
Bohang
B
Chen
Zhimin
Z
Wang
Dongming
D
Xing
Yi
Y
Ding
Lijun
L
Du
D000485:Allergens / Q000032:analysis* / Q000276:immunology*; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000737:chemistry*; D005786:Gene Expression Regulation / Q000276:immunology; D056945:Hep G2 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics*; D007267:Injections; D008516:Medicine, Chinese Traditional / Q000592:standards; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011786:Quality Control
D000485:Allergens; D004365:Drugs, Chinese Herbal; D007074:Immunoglobulin G