人结肠粘膜Mr40KD蛋白的提取、纯化及免疫学特性鉴定

目的 探索非特异性溃疡性结肠炎(UC)血清学诊断方法。方法 采用DE-52离子交换层折及cNBr-Sepharose 4B亲和层析技术从人结肠粘膜组织中提取一种蛋白组分，经10％SDS-PAGE、IEF、CIE及Westem blot技术，对该蛋白分子量、等电点及免疫学等进行鉴定。结果 产物PI为5．8，分子量为40KD，免疫学鉴定发现其主要与UC患者血清起反应。结论 人结肠40KD蛋白可能是UC的一种自身抗原，血清中ACA可以作为UC的一个特异性指标。

Extraction and Purification of Mr40KD protein from human colon tissue and it's identification on immunological property

ve To study the diagnose of the patients whth idiopathic ulcerative colitis(UC) in sera. Method The protein was purified from colon tissue using DE-52 ion exchange chromatography and CNBr-sepharose 4B affinity-column chromatography. The protein was electrophoresed in 10% SDS-PAGE and in 5% PAA gel for IEF then stained with coomassie Brilliout Blue. Using western blot the protein from colon tissue extract was transferred to nitrocellulose sheets from SDS-PAA gels, and detected by immunological procedures. Results Its PI was 5.8, with a apparent molecular weight of 40KD. By immunological procedures, it was found that this antigen tends to respond pre-dominantly to the specific antibody of sera in the patients with UC. Conclusions The human colonic 40KD protein may be a auto-antigen in the sera of many patients with symptomatic UC. The ACA (anti-colon antibodies) of sera may be a valuable and specific parameter in diagnosing UC.