Assessment of chemokine profiles in human skin biopsies by an immunoaffinity capillary electrophoresis chip.
Assessment of chemokine profiles in human skin biopsies by an immunoaffinity capillary electrophoresis chip.
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DOI：
10.1016/j.ymeth.2011.12.003
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发表时间：
2012-02
期刊：
影响因子：
4.8
通讯作者：
Phillips, Terry M.
中科院分区：
文献类型：
--
作者：
Kalish, Heather;Phillips, Terry M.
特应性皮炎是一种皮肤状况，导致皮肤皮疹暴露于环境因素。对特应性皮炎患者的皮肤活检进行显微解剖，并使用基于微芯片的免疫亲和CE系统分析CXCL1、CXCL5和CXCL8以及CCL1、CCL3和CCL5趋化因子的存在。用固定抗体的一次性免疫亲和盘从匀浆的皮肤样品中捕获CXC和CC趋化因子。然后用AlexaFluor 633标记捕获的分析物，从磁盘中洗脱并通过CE分离。用激光诱导荧光法对标记的趋化因子进行鉴定和定量。总分析时间小于40 min，包括活检显微解剖、样品分析前准备和ICE-CHIP分析，用时小于10 min，测定间和测定内CV均小于6.4%。基于微芯片的免疫亲和CE可以区分正常皮肤活检和炎症。中性粒细胞组织病理学浸润组CXCL1、CXCL5、CXCL8浓度升高，单核细胞和t淋巴细胞组织病理学浸润组CCL1、CCL3、CCL5浓度升高。该系统证明了从临床组织病理学样本中提取的冷冻切片中识别和定量免疫化学分析的能力。
Atopic dermatitis is a skin condition resulting in a skin rash from exposure to environmental factors. Skin biopsies taken from patients suffering from atopic dermatitis were micro-dissected and analyzed using a microchip-based immunoaffinity CE system for the presence of CXCL1, CXCL5 and CXCL8 and CCL1, CCL3 and CCL5 chemokines. Disposable immunoaffinity disks with immobilized antibodies were used to capture the CXC and CC chemokines from the homogenized skin samples. The captured analytes were then labeled with AlexaFluor 633, eluted from the disk and separated by CE. The labeled chemokines were identified and quantified by laser induced fluorescence. The total analysis time was less than 40 min, including the biopsy microdissection, pre-analysis preparation of the sample and the ICE-CHIP analysis, which took less than 10 min with inter- and intra-assay CV's below 6.4%. Microchip-based immunoaffinity CE could distinguish between normal skin biopsies and those with inflammation. Patients with neutrophil cellular infiltrates by histopathology showed increased concentrations of CXCL1, CXCL5 and CXCL8 while increases of CCL1, CCL3 and CCL5 corresponded to the patient group demonstrating monocytic and T-lymphocyte infiltration by histopathology. This system demonstrates the ability to identify and quantify immunochemical analytes in frozen sections taken from clinical histopathology samples.
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影响因子：
2.9
作者：
Demianova, Zuzana;Shimmo, Masahiko;Baumann, Marc
通讯作者：
Baumann, Marc
影响因子：
4.3
作者：
Guzman, NA
通讯作者：
Guzman, NA
DOI：
10.1016/j.ddtec.2006.03.009
发表时间：
2006-01-01
期刊：
Drug discovery today. Technologies
影响因子：
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作者：
Guzman, Norberto A;Stubbs, R John;Phillips, Terry M
通讯作者：
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影响因子：
7.4
作者：
Haes, Amanda J.;Terray, Alex;Collins, Greg E.
通讯作者：
Collins, Greg E.
影响因子：
4.1
作者：
Phillips, TM;Wellner, E
通讯作者：
Wellner, E