Concentration and Preservation of Very Low Abundance Biomarkers in Urine, such as Human Growth Hormone (hGH), by Cibacron Blue F3G-A Loaded Hydrogel Particles.
Concentration and Preservation of Very Low Abundance Biomarkers in Urine, such as Human Growth Hormone (hGH), by Cibacron Blue F3G-A Loaded Hydrogel Particles.
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DOI：
10.1007/s12274-008-8054-z
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发表时间：
2008-12
期刊：
影响因子：
9.9
通讯作者：
Luchini, Alessandra
中科院分区：
文献类型：
--
作者：
Fredolini, Claudia;Meani, Francesco;Reeder, K. Alex;Rucker, Sally;Patanarut, Alexis;Botterell, Palma J.;Bishop, Barney;Longo, Caterina;Espina, Virginia;Petricoin, Emanuel F., III;Liotta, Lance A.;Luchini, Alessandra
关键词：
尿液是用于检测疾病的诊断生物标志物的潜在来源，并且是非常有吸引力的非侵入性生物标本收集手段。尽管如此，尿液中的蛋白质组学测量是非常具有挑战性的，因为诊断生物标志物以非常低的浓度存在（通常低于常见免疫测定的灵敏度）并且可能会快速降解。用Cibacron Blue F3 G-A（CB）功能化的水凝胶纳米颗粒已被应用于解决尿液生物标志物测量的这些挑战。我们选择了一个最困难的低丰度，但医学相关，激素在尿液中：人类生长激素（hGH）。血清中hGH的正常范围为1 - 10 ng/mL，但怀疑尿液浓度低1000倍，远低于敏感临床hGH免疫测定的检测限（50 pg/mL）。我们证明，CB颗粒可以捕获，保存和浓缩hGH在尿液中的生理盐和尿素浓度，使hGH可以测量的线性范围内的临床免疫测定。从合成和人尿液中捕获重组和尸体hGH，浓缩并采用Immulite荧光免疫测定法进行测量。将低于0.05 ng/mL（Immulite检测限）的hGH值浓缩至2 ng/mL，尿量为1 mL。使用10 mL尿液进行的剂量反应研究表明，颗粒载体中hGH的浓度与起始溶液中hGH的浓度呈线性相关，并且所有hGH均从溶液中去除。因此，如果起始尿量为100 mL，则检测限为0.1 pg/mL。研究了血清hGH浓度为1.34 ng/mL的健康供体的尿液，以检测内源性hGH。从33 mL体积开始，颗粒洗脱液的hGH浓度为58 pg/mL，估计尿液中hGH的初始浓度为0.175 pg/mL。这里描述的纳米技术似乎具有所需的精密度，准确度和灵敏度，以支持尿液hGH水平的大规模临床研究。
Urine is a potential source of diagnostic biomarkers for detection of diseases, and is a very attractive means of non-invasive biospecimen collection. Nonetheless, proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration (usually below the sensitivity of common immunoassays) and may be subject to rapid degradation. Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A (CB) have been applied to address these challenges for urine biomarker measurement. We chose one of the most difficult low abundance, but medically relevant, hormones in the urine: human growth hormone (hGH). The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less, well below the detection limit (50 pg/mL) of sensitive clinical hGH immunoassays. We demonstrate that CB particles can capture, preserve and concentrate hGH in urine at physiological salt and urea concentrations, so that hGH can be measured in the linear range of a clinical immunometric assay. Recombinant and cadaveric hGH were captured from synthetic and human urine, concentrated and measured with an Immulite chemiluminescent immunoassay. Values of hGH less than 0.05 ng/mL (the Immulite detection limit) were concentrated to 2 ng/mL, with a urine volume of 1 mL. Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution, and that all hGH was removed from solution. Thus if the starting urine volume is 100 mL, the detection limit will be 0.1 pg/mL. Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH. Starting from a volume of 33 mL, the particle eluate had an hGH concentration of 58 pg/mL, giving an estimated initial concentration of hGH in urine of 0.175 pg/mL. The nanotechnology described here appears to have the desired precision, accuracy and sensitivity to support large scale clinical studies of urine hGH levels.
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影响因子：
3.8
作者：
Lee, Hye Lin;Eom, Hyeon-Seok;Kong, Sun-Young
通讯作者：
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作者：
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影响因子：
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作者：
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DOI：
10.1007/s10847-004-5238-3
发表时间：
2005-02-01
影响因子：
2.3
作者：
Cai, WS;Yao, XX;Pan, ZX
通讯作者：
Pan, ZX