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建立并评估一种用于循环免疫复合物中抗体的通用解离技术。

Establishment and evaluation of a general dissociation technique for antibodies in circulating immune complexes.

机构信息

Faculty of Graduate Studies, Bengbu Medical College, Bengbu, China.

Department of Clinical Laboratory Science, The 117th Hospital of PLA, 14 Lingyin Road, Westlake District, Hangzhou, 310013, China.

出版信息

Clin Exp Med. 2019 Feb;19(1):65-75. doi: 10.1007/s10238-018-0523-4. Epub 2018 Aug 17.

Abstract

This study aimed to establish a general and efficient dissociation technique for detecting antibodies in circulating immune complexes (CICs) in serum and to evaluate its clinical application. CICs were efficiently separated from specimens using polyethylene glycol double-precipitation. The best conditions for anti-HBs dissociation from HBsAg-ICs were a pH of 1.80, incubation at 15 °C for 5-10 min, and detection within 10 min after neutralization. The mean dissociation rate, reproducibility, mean dissociation recovery rate and specificity of the new technique were 64.3%, < 5.97, 95.4 and 100%, respectively. They had a favourable linear relationship (r = 0.9932), and the stability of the reagents exceeded 24 months, except the CIC antibody dissociation reagent (> 12 months). Conditions for the dissociation of other CICs tested were similar, but there were differences in the rate of antibody dissociation. Different HBV-M patterns had significantly different levels and rates of antibody dissociation from HBsAg-IC (P < 0.05), and the detection rates of the corresponding antibodies in HCV, core-anti-HCV core antibody (HCV-ICs), HIV P24-anti-HIV P24 antibody (HIV-ICs), insulin-anti-insulin antibody (INS-ICs) and thyroid globulin-anti-thyroid globulin antibody CICs (TG-ICs) were 34.8, 66.7, 20 and 14.3%, respectively. These data suggest that our CIC antibody dissociation technique is a good general pretreatment technique for the detection of antibodies after the precipitation, separation and dissociation of multiple CICs.

摘要

本研究旨在建立一种通用且高效的分离技术,用于检测血清中循环免疫复合物(CIC)中的抗体,并评估其临床应用。采用聚乙二醇双重沉淀法从标本中高效分离 CIC。从 HBsAg-IC 中分离抗-HBs 的最佳条件为 pH 为 1.80,在 15°C 孵育 5-10 分钟,中和后 10 分钟内检测。新方法的平均解离率、重现性、平均解离回收率和特异性分别为 64.3%、<5.97、95.4%和 100%。它们具有良好的线性关系(r=0.9932),试剂的稳定性超过 24 个月,除 CIC 抗体解离试剂(>12 个月)外。测试的其他 CIC 解离条件相似,但抗体解离率存在差异。不同的 HBV-M 模式对 HBsAg-IC 中抗体的解离水平和速度有显著差异(P<0.05),相应抗体在 HCV、核心抗体抗 HCV 核心抗体(HCV-ICs)、HIV P24-抗 HIV P24 抗体(HIV-ICs)、胰岛素-抗胰岛素抗体(INS-ICs)和甲状腺球蛋白-抗甲状腺球蛋白抗体 CICs(TG-ICs)中的检测率分别为 34.8%、66.7%、20%和 14.3%。这些数据表明,我们的 CIC 抗体解离技术是一种很好的通用预处理技术,可用于检测多种 CIC 沉淀、分离和解离后的抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42c1/6394588/cd4e71abdede/10238_2018_523_Fig1_HTML.jpg

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