ISOLATION AND STRUCTURE ANALYSIS OF SOLUBLE TRANSFERIN RECEPTOR FOR INTERNATIONAL ASSAY SYSTEM

国际检测系统可溶性转铁蛋白受体的分离和结构分析

基本信息

批准号：
11557071
负责人：
KOHGO Yutaka
金额：
$ 3.97万
依托单位：
ASAHIKAWA MEDICAL COLLEGE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

The concentration of soluble transferrin receptors (sTfR) is widely recognized as a good serum marker reflecting the total erythropoirsis and intracellular iron levels of erythron. While, several ELISA kits to quantify serum sTfR have been developed, the values of scram sTfR are different among these EIJSA kite. These differences are mainly caused by the standard of sTfR and the mAbs to be used. Therefore, it is important to clarify the molecular form of native sTfR complex in serum for standardization of the sTfR assay. We elucidated the structure of purified sTfR complex obtained from human serum without a detergent and dissociation procedure by means of affinity chromolography using monoclonal anti-TfR antibody. The sTfR complex was considered to be composed of two sTfR and one transferrin (Tf) and the ratio of sTfR and Tf in the complex would change depend on the serum iron concentration and iron saturation of Tf. We then developed three monoclonal antibodies against different epitopes of TfR and investigated the antibody-reactivity against sTfR complex derived from hematopoietic disease (iron deficiency anemia: IDA) and non-hematopoielic disease (chronic hepatitis: CH and rheumatoid arthritis: RA) using the two sets of ELISA. The antigenesity of sTfR in CH and RA was different from that in IDA. Because the sTfR increased in CH and RA was derived from non-erythroid cell, sTfR structure was suggested to be different between erythroid and non-erythroid cells. These results suggest thatthe sTfR derived from erythroid cell has a specific structure and the establishment of sTfR assay system against this structure will be useful to evaluate the true erythropoiesis and iro status of erythron.

可溶性转铁蛋白受体（STFR）的浓度被广泛认为是良好的血清标记物，反映了总红细胞生成和细胞内铁水平。尽管已经开发了几个用于量化血清STFR的ELISA试剂盒，但SCRAM STFR的值在这些EIJSA风筝中是不同的。这些差异主要是由STFR和MAB的标准引起的。因此，重要的是要阐明血清中天然STFR复合物的分子形式，以进行STFR分析的标准化。我们阐明了使用单克隆抗TFR抗体的亲和力染色体术，阐明了从人血清中获得的纯化的STFR复合物的结构。 STFR复合物被认为由两个STFR和一个转铁蛋白（TF）组成，并且在复合物中的STFR和TF的比率将改变取决于TF的血清铁浓度和铁饱和度。然后，我们开发了针对TFR不同表位的三种单克隆抗体，并研究了针对由造血疾病（铁缺乏症贫血：IDA）和非肺炎肝疾病（慢性肝炎：CH和Rheumatoid Chranitis：ra）衍生出的STFR复合物的抗体反应性。 Elisa集。 CH和RA中STFR的抗原性与IDA中的抗原性不同。由于CH和RA中STFR的增加是从非果皮细胞衍生而来的，因此建议STFR结构在红细胞和非果蝇细胞之间有所不同。这些结果表明，源自红细胞细胞的STFR具有特定的结构，并且针对该结构的STFR分析系统的建立对于评估Erythron的真实红细胞生成和IRO状态将很有用。