Preparation of human transferrin and verification of the antigenic activity
10.3969/j.issn.1673-4130.2018.07.004
- VernacularTitle:人转铁蛋白纯化制备及其抗原活性验证
- Author:
Qiang WU
1
;
Dinggang SHI
;
Yusheng ZHANG
Author Information
1. 美康生物科技股份有限公司
- Keywords:
transferrin;
ammonium sulfate precipitation;
anion exchange chromatography;
antigen reactivity;
immunogenicity
- From:
International Journal of Laboratory Medicine
2018;39(7):779-783
- CountryChina
- Language:Chinese
-
Abstract:
Objective Non-affinity methods were used to purify transferrin(TRF)with high purity from human serum,and the immunogenicity of TRF was evaluated by immunizing New Zealand rabbits.Methods Firstly,TRF was extracted from serum by precipitation with ammonium sulfate and then purified by two-step anion exchange chromatography.Results SDS-PAGE purity of the prepared TRF was similar with the control pure product,and the HPLC purity reached to 96.8% with a yield of 78.6%.For the same batch of TRF sam-ple,the ratio between the activity concentration determinated using TRF kit(immunoturbidimetry method) and the protein concentration determinated using uv-spectrophotometric method was about 0.95,which indica-ted that the prepared TRF for antigen could react well with the TRF antibody included in the TRF kit.Final-ly,New Zealand rabbits were immunized using the purified TRF,and the titer of the rabbit anti-serum could reach 1:128 000 after four time immunization,which also indicated that the prepared TRF had good immuno-genicity.Conclusion The TRF with high purity had good antigen reactivity and immunogenicity to prepare anti-T RF antibody by immunizing rabbits,w hich could provide qualified materials for the production of T RF kit(immunoturbidimetry method).
