Core-shell hydrogel particles harvest, concentrate and preserve labile low abundance biomarkers.
Core-shell hydrogel particles harvest, concentrate and preserve labile low abundance biomarkers.
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核壳水凝胶颗粒收获、浓缩和保存不稳定的低丰度生物标志物。
DOI：
10.1371/journal.pone.0004763
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Luchini, Alessandra
中科院分区：
文献类型：
--
作者：
Longo, Caterina;Patanarut, Alexis;George, Tony;Bishop, Barney;Zhou, Weidong;Fredolini, Claudia;Ross, Mark M.;Espina, Virginia;Pellacani, Giovanni;Petricoin, Emanuel F., III;Liotta, Lance A.;Luchini, Alessandra
血液蛋白质组被认为是早期疾病检测的生物标志物的丰富来源。然而，三个主要挑战阻碍了生物标志物的发现：a）候选生物标志物在血液中以极低的浓度存在; B）高丰度的驻留蛋白质如白蛋白掩盖了罕见的生物标志物; c）生物标志物被内源性和外源性蛋白酶快速降解。研究了用基于N-异丙基丙烯酰胺的核（365 nm）-壳（167 nm）产生并用基于电荷的诱饵（丙烯酸）官能化的水凝胶纳米颗粒，作为一种在一个步骤中解决所有这些生物标志物发现问题的技术。这些收获核-壳纳米颗粒被设计为在溶液中同时进行尺寸排阻和亲和色谱。选择血小板衍生生长因子（PDGF）作为模型，PDGF是一种临床相关、高度不稳定且丰度极低的生物标志物。加入人血清中的PDGF在几分钟内就被颗粒完全从其载体蛋白白蛋白中分离出来，浓缩并完全保存下来。完全保护颗粒隔离的PDGF免受外源性添加的胰蛋白酶降解。当将纳米颗粒以不可检测的水平（小于20皮克/mL）添加到1 mL的PDGF稀溶液中时，从颗粒的聚合物基质中释放的PDGF浓度在ELISA和质谱法的检测范围内增加。除PDGF外，还证实了一系列额外的非常低丰度和非常不稳定的细胞因子的螯合和保护免于降解。我们设想将收获核-壳纳米颗粒应用于全血，用于在静脉穿刺时浓缩和立即保存低丰度和不稳定的分析物。
The blood proteome is thought to represent a rich source of biomarkers for early stage disease detection. Nevertheless, three major challenges have hindered biomarker discovery: a) candidate biomarkers exist at extremely low concentrations in blood; b) high abundance resident proteins such as albumin mask the rare biomarkers; c) biomarkers are rapidly degraded by endogenous and exogenous proteinases. Hydrogel nanoparticles created with a N-isopropylacrylamide based core (365 nm)-shell (167 nm) and functionalized with a charged based bait (acrylic acid) were studied as a technology for addressing all these biomarker discovery problems, in one step, in solution. These harvesting core-shell nanoparticles are designed to simultaneously conduct size exclusion and affinity chromatography in solution. Platelet derived growth factor (PDGF), a clinically relevant, highly labile, and very low abundance biomarker, was chosen as a model. PDGF, spiked in human serum, was completely sequestered from its carrier protein albumin, concentrated, and fully preserved, within minutes by the particles. Particle sequestered PDGF was fully protected from exogenously added tryptic degradation. When the nanoparticles were added to a 1 mL dilute solution of PDGF at non detectable levels (less than 20 picograms per mL) the concentration of the PDGF released from the polymeric matrix of the particles increased within the detection range of ELISA and mass spectrometry. Beyond PDGF, the sequestration and protection from degradation for a series of additional very low abundance and very labile cytokines were verified. We envision the application of harvesting core-shell nanoparticles to whole blood for concentration and immediate preservation of low abundance and labile analytes at the time of venipuncture.
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影响因子：
3.5
作者：
Ayache, Saleh;Panelli, Monica;Stroncek, David F.
通讯作者：
Stroncek, David F.
影响因子：
3.5
作者：
Baxter, EJ;Hochhaus, A;Cross, NCP
通讯作者：
Cross, NCP
影响因子：
9.9
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
通讯作者：
Luchini, Alessandra
通过灵活的对接算法实现类固醇与环糊精的双峰络合
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
影响因子：
6.2
作者：
Frederick, M J;Clayman, G L
通讯作者：
Clayman, G L