Synthesis of artificial diethylstilbestrol antigen for preparation of polyclonal antibodies.
- Author:
Shujie ZHANG
1
;
Yajing LEI
;
Xiaoqian XU
;
Weixing SHI
;
Shuqing CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies; immunology; Antigens; chemistry; immunology; Diethylstilbestrol; analogs & derivatives; immunology; Enzyme-Linked Immunosorbent Assay; methods; Male; Rabbits
- From: Journal of Zhejiang University. Medical sciences 2013;42(1):25-31
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo synthesize artificial diethylstilbestrol (DES) antigen and to prepare DES polyclonal antibody with high titer and sensitivity.
METHODSThe derivative of DES (DES-HS) was synthesized from diethylstilbestrol, ethyl bromoacetate,bovine serum albumin (BSA) and chicken ovalbumin (OVA) with the nucleophilic substitution reaction; the compound was identified by electrospray ionization mass spectrometry(ESI-MS). The DES-HS and the carrier proteins (BSA, OVA) were cross-linked to prepare the artificial antigen; the UV absorption spectrophotometry and high performance liquid chromatography (HPLC) were used to identify the prepared artificial antigen. The rabbits were immunized with the DES artificial antigen to prepare the DES polyclonal antibodies.
RESULTSThe DES-HS was synthesized. The DES artificial antigen was prepared successfully with a coupling rate of 22:1. The DES polyclonal antibodies with a titer of 1:25 600 and IC50 of 10.81 ng/ml were prepared with DES artificial antigen.
CONCLUSIONA set of methods to synthesize DES artificial antigen and to prepare the DES polyclonal antibodies has been developed successfully.
