Synthesis and identification of artificial antigen of hyodeoxycholic acid
10.7501/j.issn.0253-2670.2014.23.006
- Author:
Yan ZHAO
1
Author Information
1. School of Preclinical Medicine, Beijing University of Chinese Medicine
- Publication Type:Journal Article
- Keywords:
Artificial antigen;
ELISA;
Hyodeoxycholic acid;
Mixed anhydride method;
Synthesis;
Thin layer chromatography
- From:
Chinese Traditional and Herbal Drugs
2014;45(23):3387-3391
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To synthesize and identify the hyodeoxycholic acid (HDCA) artificial antigen and to lay a foundation for the preparation of its specific monoclonal antibody, establishment of immunoassay method, and application in the pharmacological researches. Methods: The HDCA artificial immunogen (HDCA-BSA) and coating antigen (HDCA-OVA) were synthesized by mixed anhydride method and then examined by TLC method. BALB/c mice were immunized with the emulgator containing the prepared HDCA-BSA conjugate and Freund's adjuvant by the back sc multipoint injection for three times following with a booster immunization by the prepared HDCA-BSA conjugate alone. Then the blood of mice was obtained from tail vein 24 h after the booster, and the sera were isolated for the titer and specificity test by ELISA. Results: According to TLC, HDCA was successfully conjugated to BSA. The antibody against HDCA obtained from immunized-mice could bind to HDCA and the titer was over 1:8 000. As shown in the competitive inhibition curves, the antibodies in the serum were specifically bound to HDCA without cross-relativities with glycyrrhizic acid, baicalin, geniposide, etc, except cholic acid (90%). Conclusion: The HDCA-BSA is successfully synthesized, which could be used for the preparation of anti-HDCA monoclonal antibody and the research on pharmacological targets of HDCA.
