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多序列表面等离子体共振分析恶性和良性前列腺疾病患者血清中触珠蛋白-凝集素复合物。

Multi-sequential surface plasmon resonance analysis of haptoglobin-lectin complex in sera of patients with malignant and benign prostate diseases.

机构信息

Laboratory of Proteomics and Biomolecular Science, BioMedical Research Center, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.

出版信息

Anal Biochem. 2011 Dec 15;419(2):241-9. doi: 10.1016/j.ab.2011.08.029. Epub 2011 Aug 23.

Abstract

Screening for prostate cancer remains unsatisfactory. Recent studies have examined the cancer diagnostic/prognostic values of various acute phase proteins, such as haptoglobin. We describe here a novel method of surface plasmon resonance (SPR) based on multi-sequential analysis with SNA-1, AAL, and PHA-L(4) lectin, to estimate the glycosylation status of haptoglobin in sera of patients with prostate cancer (n=15), benign prostate disease (BPD) including benign prostatic hypertrophy (n=20), and normal subjects (n=11). The SPR-based analysis involves the use of anti-haptoglobin as ligand and dilution of the analyte to 1400-fold and filtration, followed by detection of the sugar chain by lectin solution. The normalized RU of lectin to haptoglobin represents the binding amount of lectin divided by that of haptoglobin. The normalized RU by SNA-1 of the prostate cancer group was significantly higher than those of the control and BPD group. SNA-1 detected NeuAcα2,6 in a biantennary sugar chain, whose content was the highest among the major glycoproteins in serum. Serum samples diluted about 7000-fold were subjected to microanalysis at 10 ng/μl and 10 μl/min for 4 min. The combination of SNA-1 and haptoglobin by SPR multi-sequential analysis offered the most accurate diagnosis of prostate cancer without any modification of serum glycoproteins.

摘要

前列腺癌的筛查仍然不尽如人意。最近的研究已经研究了各种急性期蛋白(如触珠蛋白)在癌症诊断/预后方面的价值。我们在这里描述了一种基于表面等离子体共振(SPR)的新方法,该方法基于 SNA-1、AAL 和 PHA-L(4) 凝集素的多序列分析,用于估计前列腺癌患者(n=15)、良性前列腺疾病(BPD)包括良性前列腺增生(n=20)和正常受试者(n=11)血清中触珠蛋白的糖基化状态。基于 SPR 的分析涉及使用抗触珠蛋白作为配体,并将分析物稀释 1400 倍并过滤,然后用凝集素溶液检测糖链。凝集素与触珠蛋白的归一化 RU 代表了凝集素与触珠蛋白的结合量。前列腺癌组的 SNA-1 归一化 RU 明显高于对照组和 BPD 组。SNA-1 检测到双天线糖链中的 NeuAcα2,6,其含量在血清中主要糖蛋白中最高。将血清样品稀释约 7000 倍,以 10ng/μl 和 10μl/min 的速度进行 4 分钟的微分析。SPR 多序列分析中 SNA-1 和触珠蛋白的结合提供了最准确的前列腺癌诊断,而无需对血清糖蛋白进行任何修饰。

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