The Research Of Sreening Carrier Protein Of Sensitinogens Of SHL Injection

Objective:Taking Shuanghuanglian injection as the object of study, carrier protein of Shuanghuanglian injection allergen group were screened through combining proteomics, enzyme-linked immunosorbent assay, HPLC, Shuanghuanglian intervention, and using mass spectrometry to identify the carrier protein, it can fully elucidate a sensing mechanism caused by Shuanghuanglian Injection, and provide ideas for screening carrier protein of other TCMI sensitization component.Methods:Firstly, sensitized serum was received by animal sensitization, the level of serum IgE was determinated by ELISA kit, Study on differential proteomics of sensitized serum and blank serum through two-dimensional electrophoresis, a preliminary study on the effect of Shuanghuanglian Injection on serum protein, lay the foundation for the next step of Screening of carrier protein; Separating the serum IgE and IgG, and comparing the differents of immune fingerprints of oral and injected shuanghuanglian allergenic ingredients of Shuanghuanglian injection was screened by enzyme immunoassay method combined with HPLC; Carrier protein of Shuanghuanglian injection were screened by Shuanghuanglian injection, chlorogenic acid, baicalin, which verifies the carrier protein screened, but also validate the screening way. Through query the database of the Mascot,5proteins were identified success sfully, they are TRYP_PIG,K2C1_PANTR,TRY1_RAT, WNT7A_AOTTR, CHIT_MANSE.Conclusion:Baicalin and Chlorogenic Acid of Shuanghuanglian injection are the sensitization component, its carrier protein may be TRYP_PIG, K2C1_PANTR, TRY1_RAT, WNT7A_AOTTR, CHIT_MANSE. The sensitization mechanism might also be associated with supramolecular mechanism.