CN1697975A - 包含纳米结构的分析或分离用装置、制备方法及其应用 - Google Patents
包含纳米结构的分析或分离用装置、制备方法及其应用 Download PDFInfo
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- CN1697975A CN1697975A CNA2004800006534A CN200480000653A CN1697975A CN 1697975 A CN1697975 A CN 1697975A CN A2004800006534 A CNA2004800006534 A CN A2004800006534A CN 200480000653 A CN200480000653 A CN 200480000653A CN 1697975 A CN1697975 A CN 1697975A
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Abstract
Description
Claims (60)
- 权利要求1、 一种用于分离或 /和分析的纳米结构载体, 其包含纳米结构区, 所述纳 米结构区包含常规固相载体及其上固定的纳米结构, 所述纳米结构包含非定向 排列的纳米结构单元, 所述纳米结构单元包括凸出高度大于 3 nm、 且凸出半高 处至少一维尺寸在 1一 500 纖、 优选 1― 1 OOnm之间的纳米凸体, 且所述纳米凸 体在所述纳米结构区中的分布密度大于 1个 / μ m2、 优选大于 10个 / μ m2。
- 2、 根据权利要求 1所述的纳米结构载体, 其中所述纳米结构区中纳米结构 覆盖率大于 20%、 优选大于 30%。
- 3、根据权利要求 1或 2所述的纳米结构载体,其中所述纳米结构区的表面积 提高率大于 200%、 优选大于 300%、 更优选大于 400%。
- 4、根据权利要求 1一 3之一所述的纳米结构载体,其中所述纳米结构区的吸 附能力提高率大于 115 %、 优选大于 130%、 更优选大于 150%。
- 5、根据权利要求 1一 4之一所述的纳米结构载体,其中所述纳米结构区的吸 附量衰减速率比小于 90%、 优选小于 80%。
- 6、根据权利要求 1一 5之一所述的纳米结构载体,其中所述纳米结构单元由 纳米粒子形成, 所述纳米粒子其粒径尺寸 1一 500nm、 优选 1一 100nm、 更优选 1 — 50nm。
- 7、 根据权利要求 6所述的纳米结构载体, 其中所述纳米粒子包括无机纳米 粒子或 /和有机纳米粒子及它们的衍生物, 所述无机纳米粒子包括非磁性无机纳 米粒子和磁性无机纳米粒子, 所述非磁性无机纳米粒子包括非磁性金属纳米粒 子和非磁性非金属纳米粒子, 所述衍生物包括结合有表面基团或 /和包被有机物 的衍生物。
- 8、 根据权利要求 7所述的纳米结构载体, 其中所述无机非磁性非金属纳米 粒子包括氧化硅粒子、 氧化钛粒子、 氧化铝粒子在内的氧化物纳米粒子。
- 9、 根据权利要求 7所述的纳米结构载体, 其中所述非磁性金属纳米粒子包 括金粒子、 钒粒子、 铅粒子。
- 10、 根据权利要求 1一 9之一所述的纳米结构载体, 其中所述常规固相载体 包括由以下之一组或多组材料或其衍生物制成的尺寸大于 lOOOnm的固相载体: 玻璃、 硅片、 硅胶、 陶瓷、 金属氧化物、 金属、 聚合物材料及它们的复合物, 所述衍生物包括结合有表面基团或 /和包被有机物的衍生物。
- 11、根据权利要求 8— 10之一所述的纳米结构载体,其中所述表面基团包括 下述一种或多种基团: 氨基、 醛基、 环氧基、 氨基肼、 二乙氨基乙基、 二乙基 一 (2—羟丙基)氨乙基、 羧甲基、磺酸丙基、 巯乙基吡啶基、 硅氧垸基、硫醇 基、垸基; 所述包被有机物包括下述一种或多种有机物:包括聚乙烯吡咯垸酮、 吐温类表面活性剂在内的表面活性剂, 包括聚氨基酸在内的聚电解质, 包括聚 硅氧烷在内的亲油有机物, 包括葡聚糖衍生物、琼脂糖衍生物、纤维素衍生物、 聚丙烯酰胺在内的离子交换聚合物, 和包括肝素钠、 抗原、 抗体在内的亲和物 质。
- 12、根据权利要求 1一 11之一所述的纳米结构载体,其中所述纳米结构载体 包括: 纳米粒子 /片基复合物、 纳米粒子 /微米粒子复合物或纳米粒子 /微米粒子 / 片基复合物。
- 13、根据权利要求 1一 12之一所述的纳米结构载体,其为下述组之一: 分析 芯片纳米结构片基、 用以制作酶标板的纳米结构微孔板、 平面层析纳米结构片 基、 纳米结构分离固定相、 及包括纳米结构层析凝胶在内的纳米结构分离固定 相。14、一种制备如权利要求 1一 13之一所述的纳米结构载体的方法,其包括将 所述纳米粒子的悬浮液与所述常规固相载体接触并进行固定化反应, 且固定化 反应时所述悬浮液中的纳米粒子重量 /体积浓度(g/ml)为五百分之一至十五万 分之一, 或纳米粒子摩尔浓度为 0.12— 37.4 nM。15、根据权利要求 14所述的方法,其中所述纳米粒子重量 /体积浓度(g/ml) 为五千分之一至十五万分之一, 或纳米粒子摩尔浓度为 0.12— 3.74 nM。16、根据权利要求 14所述的方法,其中所述纳米粒子重量 /体积浓度(g/ml) 为二万分之一至六万分之一, 或纳米粒子摩尔浓度为 0.31— 0.93 nM。
- 17、 根据权利要求 14一 16之一所述的方法, 其还包括下述一种或多种功能 化处理:(a) 先将所述纳米粒子或 /和所述常规固相载体功能化, 再进行所述接触 和固化;(b) 先进行所述接触和固化, 再进行所述纳米粒子或 /和所述常规固相载 体的功能化;(c) 将所述纳米粒子或 /和所述常规固相载体功能化, 再进行所述接触和 固化, 然后再进行所述纳米粒子或 /和所述常规固相载体的功能化;其中: 所述功能化处理包括引入所述表面基团或 /和包被有机物。
- 18、 根据权利要求 14一 17之一所述的方法, 其还包括对所述固定化反应后 的产物进行化学交联处理。
- 19、 根据权利要求 14一 18之一所述的方法, 其还包括对所述固定化反应后 的产物进行热处理, 所述热处理包括在 30°C以上、 优选 37°C以上、 更优选 40°C 以上但固相载体的软化温度以下加温然后冷却。
- 20、 一种用于分离或 /和分析反应器的纳米结构活性载体, 其包含纳米结构 活性区, 所述纳米结构活性区包含常规固相载体及其上固定的活性纳米结构, 所述活性纳米结构包含非定向排列的纳米结构单元及其上固定的活性试剂, 所 述纳米结构单元包括凸出高度大于 3 nm、 且凸出半高处至少一维尺寸在 1一 500 nm、 优选 1一 lOOrnn之间的纳米凸体, 且所述纳米凸体在所述纳米结构区中的 分布密度大于 1个 / μ πι<sup>2</sup>、 优选大于 10个 / μ ιη<sup>2</sup>。
- 21、 根据权利要求 20所述的纳米结构活性载体, 其中所述纳米结构活性区 中纳米结构覆盖率大于 30%、 优选大于 75 %。
- 22、 根据权利要求 20或 21所述的纳米结构活性载体, 其中所述纳米结构活 性区的表面积提高率大于 200%、 优选大于 400%、 更优选大于 600%, 或 /和吸 附能力提高率大于 115 %、 优选大于 130%、 更优选大于 150%。
- 23、 根据权利要求 20— 22之一所述的纳米结构活性载体, 其中所述纳米结 构活性区的灵敏度提高率大于 120%、 优选大于 150%、 更优选大于 200%。
- 24、 根据权利要求 20— 23之一所述的纳米结构活性载体, 其中所述纳米结 构活性区的识别信号衰减速率比小于 90%、优选小于 70%, 或 /和吸附量衰减速 率比小于 90%、 优选小于 80%。
- 25、根据权利要求 20— 24之一所述的纳米结构活性载体, 其中所述分离或 / 和分析的目标物包括多肽或 /和与多肽相互作用的药物。
- 26、根据权利要求 20— 25之一所述的纳米结构活性载体, 其中所述分离或 / 和分析的目标物包括核酸或 /和与核酸相互作用的药物。
- 27、 根据权利要求 20— 26之一所述的纳米结构活性载体, 其为活性纳米粒 子与固相载体的复合物, 其中: 所述活性纳米粒子包含纳米粒子和固定于其上 的一种或多种所述活性试剂或任选存在的连接剂, 所述纳米粒子粒径为 1一 500nm、 优选 1一 100nm、 更优选 1一 50nm, 所述固相载体为常规固相载体或权 利要求 1一 14之一所述纳米结构载体。
- 28、 根据权利要求 27所述的纳米结构活性载体,所述纳米粒子包括权利要 求 6— 11之一所述纳米粒子,优选为所述无机非磁性非金属纳米粒子及其所述衍 生物。 29、 根据权利要求 27或 28之一所述的纳米结构活性载体, 其包括下述组之 -: 活性纳米粒子 /片基复合物、 活性纳米粒子 /微米粒子复合物、 活性纳米粒 子 /纳米结构片基复合物、 活性纳米粒子 /纳米结构微米粒子复合物、 及它们的 组合物。
- 30、 根据权利要求 20— 26之一所述的纳米结构活性载体,其为活性试剂与 权利要求 1一 13之一所述纳米结构载体的复合物。
- 31、 根据权利要求 30所述的纳米结构活性载体, 其包括下述组之一: 活性 试剂 /纳米结构片基复合物、 活性试剂 /纳米结构微米粒子复合物、 及它们的组 合物。
- 32、 根据权利要求 20— 31之一所述的纳米结构活性载体, 其包括纳米结构 分析芯片。
- 33、 根据权利要求 20— 31之一所述的纳米结构活性载体, 其包括下述组之 一: 纳米结构酶标板、 纳米结构平面层析试剂条、 及包括纳米结构层析活性凝 胶在内的纳米结构活性分离固定相。34、 一种制备如权利要求 21— 33之一所述的纳米结构活性载体的方法, 其 包括将所述活性纳米粒子的悬浮液与所述常规固相载体接触并进行固定化反 应, 而且: 固定化反应时所述悬浮液中的纳米粒子重量 /体积浓度 (g/ml) 为五 百分之一至十五万分之一, 或纳米粒子摩尔浓度为 0.12— 37.4 nM; 所述接触为 表面直径小于 0.5mm的点接触或表面直径大于 0.'5mm的面接触。35、根据权利要求 34所述的方法,其中所述纳米粒子重量 /体积浓度(g/ml) 为五千分之一至十五万分之一, 或纳米粒子摩尔浓度为 0.12— 3.74 nM。36、根据权利要求 34所述的方法,其中所述纳米粒子重量 /体积浓度(g/ml) 为二万分之一至六万分之一, 或纳米粒子摩尔浓度为 0.31— 0.93 nM。
- 37、 根据权利要求 34— 36之一所述的方法, 其还包括对所述固定化反应后 的产物进行化学交联处理。
- 38、 根据权利要求 34— 37之一所述的方法, 其还包括对所述固定化反应后 的产物进行热处理, 所述热处理包括在任选存在的活性试剂保护剂保护下在 30 度以上、 优选 37度以上但固相载体的软化温度以下加温然后冷却。
- 39、一种分析或 /和分离装置,其含有权利要求 21— 33之一所述的纳米结构 载体或 /和权利要求 1一 14之一所述的纳米结构载体。
- 40、 根据权利要求 39所述的装置, 其包括下述组之一的分析芯片: 微阵列 芯片、 微阵列一流路芯片及流路芯片, 其中所述流路包括含所述纳米结构载体 的纳米结构流路、 或 /和含所述纳米结构活性载体的纳米结构活性流路。
- 41、 根据权利要求 39所述的装置, 其包括下述组之一: 酶标板、 平面层析 试剂条、 及包括层析柱在内的含分离固定相的装置。
- 42、 一种定量或 /和定性分析标记***, 其含标记物, 且至少一种标记物为 标记标记活性试剂 /纳米结构 /分子标记物质复合物, 所述纳米结构包括纳米粒 子或 /和由纳米粒子组成的结构, 且所述纳米粒子为粒径 1— 100 urn且本身不是 标记物质增强剂的非磁性无机非金属粒子。
- 43、 根据权利要求 42所述的标记***, 其中所述非磁性无机非金属粒子包 括权利要求 8或 9或 11所述非磁性无机非金属粒子。
- 44、 根据权利要求 42或 43所述的标记***, 其中所述分子标记物质包括下 述一种或多种物质: 荧光物质、化学发光物质、化学发光催化剂、有色金属盐、 染料和颜料。
- 45、一种定量或 /和定性分析方法, 其中包括使用权利要求 39— 41之一所述 分析或 /和分离装置来捕捉或 /和输运或 /和分离样品目标物, 及使用权利要求 42 一 44之一所述定量或 /和定性分析标记***进行标记。
- 46、一种定量或 /和定性检测试剂盒, 其包含权利要求 39— 41之一所述分析 或 /和分离装置或装置和权利要求 42— 44之一所述定量或 /和定性分析标记系 统。
- 47、根据权利要求 46所述的试剂盒, 其为用于多肽或 /和与多肽相关的药物 分析的试剂盒, 包括多肽分析芯片试剂盒、 多肽分析酶标试剂盒、 或多肽分析 平面层析试剂盒。
- 48、根据权利要求 46所述的试剂盒, 其为用于核酸或 /和与核酸相关的药物 分析的试剂盒。
- 49、一种定量或 /和定性分析方法, 其包括使用权利要求 39— 41之一所述分 析或 /和分离装置来捕捉或 /和输运或 /和分离样品目标物。
- 50、一种定量或 /和定性检测试剂盒, 其包含权利要求 39— 41之一所述分析 或 /和分离装置或装置。51、根据权利要求 50所述的试剂盒, 其为用于多肽或 /和与多肽相关的药物 分析的.试剂盒, 包括多肽分析芯片试剂盒、 多肽分析酶标试剂盒、 或多肽分析 平面层析试剂盒。
- 52、根据权利要求 50所述的试剂盒, 其为用于核酸或 /和与核酸相关的药物 分析的试剂盒。 53、一种定量或 /和定性分析方法, 其包括使用权利要求 42— 44之一所述定 量或 /和定性分析标记***进行标记。
- 54、一种定量或 /和定性检测试剂盒, 其包含权利要求 42— 44之一所述定量 或 /和定性分析标记***。
- 55、根据权利要求 54所述的试剂盒, 其为用于多肽或 /和与多肽相关的药物 分析的试剂盒, 包括多肽分析芯片试剂盒、 多肽分析酶标试剂盒、 或多肽分析 平面层析试剂盒。
- 56、根据权利要求 54所述的试剂盒, 其为用于核酸或 /和与核酸相关的药物 分析的试剂盒。
- 57、 一种芯片检测方法, 其至少包括以下步骤:(a)准备样品、 芯片及芯片标记***, 所述样品可能含有目标物, 所述芯 片包含固定化活性试剂阵列, 所述芯片标记***包含标记物、 着色剂、 和任选 存在的着色控制剂及着色定型剂, 且所述标记物包含标记活性试剂 /纳米结构 / 催化剂复合物, 其中所述纳米结构包括纳米粒子或 /和由纳米粒子组成的结构, 所述纳米粒子为粒径 1一 100 nm的无机纳米粒子或 /和有机纳米粒子或 /和它们 的衍生物, 所述无机纳米粒子包括非磁性无机纳米粒子和磁性无机纳米粒子, 所述非磁性无机纳米粒子包括非磁性金属纳米粒子和非磁性非金属纳米粒子, 所述衍生物包括结合有表面基团或 /和包被有机物的衍生物;(b)用所述固定化活性试剂固定所述目标物,然后通过所述目标物与所述 标记物中的标记活性试剂反应,固定所述标记物中的纳米结构及其上的催化剂; 或(c)使所述目标物结合在所述全部或部分标记物上,然后通过所述目标物 与所述固定化活性试剂反应, 固定所述标记物中的纳米结构及其上的催化剂;(d)加入所述着色剂并在所述催化剂作用下、在所述纳米结构上而不是在 所述固定化活性试剂与所述目标物的结合处进行着色反应, 所述可见光着色反 应中还包括任选地使用着色控制剂及定型剂;(e) 对步骤 (d) 中着色反应的结果、 及步骤 (b)或 (c) 中着色剂的着 色结果进行检测、 分析。
- 58、 根据权利要求 57所述的方法, 所述标记物中还含有染色剂, 在所述着 色反应前、 反应中、 或 /和反应后还进行染色反应, 其中所述染色剂存在于下述 之一种或多种标记物中: 标记活性试剂 /纳米结构 /催化剂 /染色剂复合物、 标记 活性试剂 /纳米结构 /染色剂复合物、 及标记活性试剂 /染色剂复合物, 其中所述 纳米结构为权利要求 57所述纳米结构。
- 59、 根据权利要求 57或 58所述的方法, 其中所述标记***中的所述纳米粒 子包括权利要求 6— 11之一所述纳米粒子。
- 60、 根据权利要求 57或 58所述的方法, 其中所述催化剂包括金属化合物还 原反应的金催化剂或 /和酶催化剂, 所述染色剂包括染料, 所述着色剂包括金属 化合物。
- 61、 一种芯片检测方法, 其至少包括以下步骤:(a)准备样品、 芯片及芯片标记***, 所述样品可能含有目标物, 所述芯 片包含固定化活性试剂阵列, 所述芯片标记***包含标记物、 着色剂、 和任选 存在的着色控制剂及着色定型剂, 且所述标记物包含活性试剂、 催化剂、 染色 剂及任选存在的纳米结构, 其中所述纳米结构、 染色剂、 催化剂、 着色剂分别 为权利要求 57— 60之一所述的芯片标记***中所述纳米结构、染色剂、催化剂、 着色剂;(b)在所述芯片中加入所述样品,在所述固定化活性试剂与可能存在的所 述目标物反应后加入所述标记物和着色剂, 并对所述反应处进行染色和着色; 或(c )将所述样品与所述全部或部分标记物混合,在所述标记活性试剂与可 能存在的所述目标物反应后加入所述芯片中, 在所述固定化活性试剂与可能存 在的所述目标物反应后加入所述着色剂, 并对芯片中所述反应处进行染色和着 色;( d) 对步骤 (b ) 或(c) 中染色和着色的结果进行检测、 分析。
- 62、 一种芯片试剂盒, 其包含标记***, 其中至少一个所述标记***为权 利要求 57— 60之一或权利要求 61所述芯片标记***。
- 63、根据权利要求 62所述的试剂盒, 其为用于多肽或 /和多肽相关药物分析 的试剂盒。
- 64、根据权利要求 62所述的试剂盒, 其为用于核酸或 /和核酸相关药物分析 的试剂盒。
- 65、 一种多肽分析的分析芯片检测方法, 其至少包括下述之一个步骤:( a)将样品与磁纳米粒子混合;(b )将样品与活性试剂 /磁纳米粒子复合物混合;( c)将样品与芯片接触并反应, 反应时可任选存在外加磁场, 所述芯片为 纳米结构芯片, 且其中所述纳米结构包含磁纳米粒子;( d) 将标记活性试剂 /磁纳米粒子 /分子标记物质复合物用于标记反应, 在 标记时可任选存在外加磁场, 所述活性试剂 /磁纳米粒子 /分子标记物质复合物 含有一种或多种分子标记物质、 一种或多种磁纳米粒子、 一种或多种活性试剂 以及任选存在的封闭剂的混合物或纯化物;其中:所述磁纳米粒子在三维空间中至少有一维为 1一 200 nm、优选 1一 100 讓、更优选 1一 50 nm,且其本身不是分子标记物质增强剂的磁粒子及其衍生物; 所述活性试剂选自以下组中能与多肽作用的物质: 多肽、 多糖、 维生素、 抗生 素、 病毒、 细胞、 及功能有机物, 所述活性试剂 /磁纳米粒子 /分子标记物质复 合物在标记时的磁纳米粒子浓度为: 纳米粒子重量 /体积浓度 (g/ml)大于三万 分之一、 或纳米粒子摩尔浓度大于 0.25nM, 优选为纳米粒子重量 /体积浓度 (g/ml)大于三千分之一、或纳米粒子摩尔浓度大于 2.4nM, 更优选为纳米粒子 重量 /体积浓度 (g/ml)大于五百分之一、 或纳米粒子摩尔浓度大于 15.0nM。
- 66、根据权利要求 65所述的方法,其中所述磁粒子选自于包括四氧化三铁、 三氧化二铁在内的铁氧体及其衍生物, 而所述衍生物包括表面含衍生基团的表 面修饰或 /和功能有机物包被衍生物。
- 67、 根据权利要求 65或 66所述的方法, 其中所述外加磁场是脉冲式的。
- 68、一种多肽分析芯片试剂盒,其包含如权利要求 65或 66所述磁纳米粒子。
- 69、 一种分离方法, 其包括使用如权利要求 39— 41之一所述分离装置。
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2004
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CN107436350B (zh) * | 2017-07-31 | 2019-05-24 | 云南沃森生物技术股份有限公司 | 一种利用酶标板和单克隆抗体进行混合物中各成分的分离方法 |
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CN100410664C (zh) | 2008-08-13 |
CN1514243A (zh) | 2004-07-21 |
US7842515B2 (en) | 2010-11-30 |
US20060057631A1 (en) | 2006-03-16 |
JP2007502998A (ja) | 2007-02-15 |
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