WO2004038413A1 - Systeme de detection de cristaux de proteine pouvant detecter simultanement plusieurs cibles - Google Patents

Systeme de detection de cristaux de proteine pouvant detecter simultanement plusieurs cibles Download PDF

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WO2004038413A1
WO2004038413A1 PCT/CN2003/000208 CN0300208W WO2004038413A1 WO 2004038413 A1 WO2004038413 A1 WO 2004038413A1 CN 0300208 W CN0300208 W CN 0300208W WO 2004038413 A1 WO2004038413 A1 WO 2004038413A1
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protein
solution
protein chip
chip
concentration
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PCT/CN2003/000208
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English (en)
French (fr)
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Gengxi Hu
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Shanghai Healthdigit Co., Ltd.
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Priority to JP2004545675A priority Critical patent/JP2006503300A/ja
Priority to US10/532,837 priority patent/US20060008793A1/en
Priority to CA002503601A priority patent/CA2503601A1/en
Priority to EP03714619A priority patent/EP1560023A4/en
Priority to AU2003227167A priority patent/AU2003227167A1/en
Publication of WO2004038413A1 publication Critical patent/WO2004038413A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00—Methods of screening libraries
    • C40B30/04—Methods of screening libraries by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54353—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals with ligand attached to the carrier via a chemical coupling agent
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6845—Methods of identifying protein-protein interactions in protein mixtures

Definitions

  • the invention relates to the field of biotechnology, and in particular, to a protein chip detection system and a preparation method for multi-index parallel detection. Background technique
  • Biochip technology is a new biological technology that emerged in the mid-1990s. It is based on a large-scale parallel analysis method of interactions between biological macromolecules (nucleic acids, proteins, etc.), and combines microelectronics, micromechanics, chemistry, physics, Computers and other fields of technology have continuously, integrated, and miniaturized the processes of sample reaction, detection, and analysis involved in life science research, and have become one of the fastest growing technologies in the field of life science research today.
  • Biochips can be widely used in agriculture, environment, food, court, military, scientific research and other fields. Not only that, because the biochip can simultaneously detect multiple disease-related genes or proteins, thereby assisting the diagnosis of genetic diseases, tumors, and infectious diseases, it also has important clinical application value.
  • Biochips are divided into three main categories: nucleic acid chips, protein chips, and chip laboratories.
  • a protein chip is a biochip that detects interactions between proteins. Most of the protein chip technologies in the current research are based on the principle of specific binding of antigens and antibodies. A variety of proteins are bound to a solid phase substrate (such as specially processed slides, organic membranes, silicon microspheres, etc.) to detect biological samples. Contains the corresponding protein that can specifically bind to it.
  • the protein chip can accurately determine physiological / pathological changes by detecting changes in the content of specific proteins in the body. In the field of clinical diagnosis, protein chips have broad application prospects.
  • the present inventor has disclosed a protein chip detection system and a preparation method capable of simultaneously detecting multiple indicators in Chinese Patent 011050233.3, and has realized clinical simultaneous detection of some proteins with high specificity but low content in body fluids. Summary of the Invention
  • the technical problem to be solved by the present invention is to improve the protein chip detection system on the basis of the original application 011050233.3 to increase the stability of the detection system and improve the detection sensitivity.
  • Chinese patent application 011050233.3 has disclosed a protein chip detection system for parallel detection of multiple indicators.
  • the detection system includes:
  • a protein chip for parallel detection of multiple indicators (2) a multi-protein mixed solution, which is prepared at a certain concentration ratio, and bears a light-emitting label, that is, a reaction solution;
  • the invention modifies the formula of the standard solution of the protein to be tested, so that different proteins will not lose activity after being mixed, the stability is increased, and the accuracy of detection is improved.
  • test protein (or target protein A) standard solution formula is 40%-60% fetal bovine serum + various high-concentration purified antigen + 0. 02-0. L% . NaN 3 or PH7. 0-7. 8 0. 05M PB (KH 2 P0 4 -Na 2 HP0 4 ) + 2-30% BSA + l. 5-2. 5% sucrose + 0.02-0. L% . NaN 3 + various high concentration purified antigens.
  • This series of known concentration and increasing concentration of the target protein A mixed solution is prepared after lyophilization.
  • Another technical problem to be solved by the present invention is to provide an improved method for preparing a protein chip in the above detection system.
  • the protein chip of the present invention includes a solid-phase carrier and proteins fixed on the carrier for multi-parameter parallel detection.
  • the protein chip uses a chip automatic spotting system to combine a variety of proteins by physical adsorption or covalent binding. Fix it on the solid-phase support in a certain arrangement order; then close the non-spotted part of the solid-phase support with a blocking solution and store it in a dry place.
  • the protein may be an antigen, an antibody, a receptor, a ligand, etc., and further refers to a protein capable of specifically binding to a disease marker protein in vivo, especially a tumor marker protein.
  • the solid phase carrier may be an inorganic substrate or an organic compound substrate.
  • the inorganic substrate includes a semiconductor wafer, a glass wafer, a microporous silicon wafer, a microporous glass wafer, etc., preferably a glass wafer.
  • the organic wafer includes acetic acid. Cellulose film, nitrocellulose film, nylon film, polypropylene film, etc.
  • the protein chip is obtained by the following technical scheme:
  • target protein A analyte proteins
  • B A-specific antigens, antibodies or receptors
  • the coating liquid formulation provided by the present invention is PB (KH 2 P0 4 -Na 2 HP0 4 ) having a pH of 7.0-8.0. 01- l% ⁇
  • the blocking solution formula provided by the present invention is: containing 1-9% BSA, 1-9% sucrose, 0.01-1%.
  • NaN 3 's PB KH 2 P0 4 -Na 2 HP0 4
  • BSA has a blocking effect
  • sucrose is an inert substance that can block the air
  • BSA and sucrose can be used as a material to support the frame structure.
  • the effect of the blocking solution The protein cannot be bound to other parts of the solid phase carrier, thereby ensuring the accuracy of the experimental data.
  • the improvement of the buffer formulation in the preparation method of the present invention reduces non-specific adsorption, reduces the background signal value, and increases the stability of the protein chip and improves the detection sensitivity.
  • FIG. 1 is a signal image of a protein chip prepared by an improved buffer formulation preparation method of the present invention.
  • Figure 2 is a signal image of a protein chip prepared by the original preparation method. detailed description:
  • the amount of solid materials (such as BSA, sucrose, tyrosine, and NaN 3 ) used is expressed by weight percentage, and the amount of used solutions (such as Tween 20, fetal bovine serum, and Procin in liquid) is expressed by volume percentage. .
  • the coating solution used was PB (KH 2 P0 4 -Na 2 HP0 4 ) having a pH of 7.8.
  • the formula of the blocking solution used was: 3% BSA, 5% sucrose, 0.5. /. . NaN 3 PB (KH 2 P0 4 -Na 2 HP0 4 ). 2. Preparation of protein chip F using the original protein chip preparation method
  • the coating solution used therein was CBS (NaHC0 3 -Na 2 C0 3 ) having a pH of 9.6.
  • the formula of the blocking solution used is: containing 0.2% Tween20, 0.1% tyrosine, 5% BSA, 4% sucrose, 0 ° / oproc l in TBS.
  • the above protein chip ⁇ F was reacted with the supporting reaction solution, washing solution, and serum samples of known concentration (the remaining serum samples were reserved for the next experiment).
  • the biochip detector was used for shooting and data analysis. The signal values are shown in Table 1:
  • the chip was stored at 4 ° C, and after 6 months, the chip was reacted with the same batch of the reaction solution, washing solution, and the remaining serum samples in the above experiment.
  • the signal values are shown in Table 2:
  • the concentrations of the standards a and b are more stable and can be stored for a longer time without losing activity.

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  • Proteomics, Peptides & Aminoacids (AREA)
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  • Bioinformatics & Computational Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
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  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
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Description

技术领域
本发明涉及生物技术领域， 具体涉及一种多指标并行检测的蛋白芯片检测系统 及制备方法。 背景技术
生物芯片技术是九十年代中期兴起的一种新型生物学技术， 它基于生物大分子 (核酸、 蛋白质等） 间相互作用的大规模并行分析方法， 并结合微电子、 微机械、 化学、 物理、 计算机等多领域的技术， 将生命科学研究中所涉及的样品反应、 检测、 分析等过程连续化、 集成化、 微型化， 已成为当今生命科学研究领域发展最快的技 术之一。
生物芯片可以广泛应用于农业、 环境、 食品、 法庭、 军事、 科研等领域。 不仅 如此， 由于生物芯片可同时检测多种疾病相关基因或蛋白， 从而对遗传病、 肿瘤、 传染性疾病进行辅助诊断， 所以它还具有重要的临床应用价值。
生物芯片主要分为三大类： 核酸芯片、 蛋白芯片和芯片实验室。
蛋白芯片是检测蛋白质之间相互作用的生物芯片。 目前研究中的蛋白芯片技术大多基 于抗原和抗体特异性结合的原理， 将多种蛋白质结合在固相基质 (如： 特殊处理的玻片、 有机膜片、 硅微球等)上， 检测生物样品所含有的可与之专一性结合的对应蛋白质。
蛋白芯片可通过发现体内特定蛋白质的含量变化，准确地对生理 /病理变化作出 判断。 在临床诊断领域， 蛋白芯片具有广阔的应用前景。
本发明人在中国专利 01105023. 3 中已公开了一种能同时检测多指标的蛋白芯 片检测系统及制备方法， 实现了对一些特异性高但在体液中含量很低蛋白的临床同 时检测。 发明内容
本发明所要解决的技术问题， 是在原申请 01105023. 3基础上， 对蛋白芯片检测 系统加以改进， 以增加检测系统的稳定性， 提高检测灵敏度。
中国专利申请 01105023. 3 中已公开了一种用于多指标并行检测的蛋白芯片检 测系统， 该检测系统包括：
( 1 )用于多指标并行检测的蛋白芯片； ( 2 ) 以一定浓度配比制成的， 并带有发光标记的多蛋白混合液即反应液；
( 3 ) 一系列已知浓度且浓度递增的被测蛋白的混合液即标准品；
( 4 ) 一种蛋白芯片的洗涤液。 .
本发明修改了被测蛋白标准品溶液的配方，使不同蛋白质混合后不会丧失活性， 增加了其稳定性， 提高了检测的准确性。
所述的已知浓度且浓度递增的被测蛋白 （或称目标蛋白 A ) 的标准品溶液配方 为 40%- 60%胎牛血清 +各种高浓度纯化抗原 +0. 02-0. l%。NaN3或 PH7. 0-7. 8 0. 05M PB (KH2P04-Na2HP04) +2-30%BSA+l. 5-2. 5%蔗糖 +0. 02-0. l%。NaN3+各种高浓度纯化抗原。 该一系列已知浓度且浓度递增的目标蛋白 A的混合液配制后冻干保存。
本发明所要解决的另一技术问题是提供一种改进的上述检测系统中的蛋白芯片 的制备方法。
本发明所述蛋白芯片包括固相载体和固定于载体上用于多指标并行检测的蛋 白， ,该蛋白芯片是利用芯片自动点样系统， 通过物理吸附或共价结合的方式， 将多 种蛋白质以一定的排列次序固定在固相载体上； 然后用封闭液将固相载体的非点样 部位封闭， 干燥保存。 蛋白质可以是抗原、 抗体、 受体、 配体等， 进一步指能与体 内疾病性标志蛋白特异结合的蛋白， 尤其是肿瘤标志性蛋白。
其中所述的固相载体可以是无机片基或有机化合物片基， 无机片基包括半导体 硅片、 玻璃片、 微孔硅片、 微孔玻璃片等， 优选玻璃片； .有机片基包括醋酸纤维素 膜、 硝酸纤维素膜， 尼龙膜， 聚丙烯膜等。
该蛋白芯片是通过下述技术方案获得的：
1、 确定多种不同的被测物蛋白 （简称目标蛋白 A )和 A特异性的抗原、 抗体或 受体等 （简称 B ), 如体内疾病标志性蛋白和能与之特异结合的多种蛋白；
1、将上述所述 B即不同的蛋白探针以一定浓度溶于包被液中，然后用芯片自动 点样系统将这些蛋白质点制在固相载体上， 点样密度 25-200 点 /cm2, 点样量 0. 1- 10ng/点；
3、 4 °C放置过夜；
4、 用封闭液将蛋白芯片封闭；
5、 干燥处理， 储存于 4 °C ;
为了使芯片上所点的多种蛋白质在发生一系列反应后产生的光信号能在相邻两 个数量级上 (最强点 /最弱点 <100) , 以便于检测仪器的检测， 事先调节了芯片上蛋白 质的浓度。 （最适点样浓度）。 本发明提供的包被液配方为 PH为 7. 0-8. 0的 PB (KH2P04-Na2HP04)。 本发明提供的封闭液配方为：含 1-9%BSA , 1-9% 蔗糖， 0. 01- l %。NaN3的 PB (KH2P04-Na2HP04)„ 其优点是： BSA都有封闭作用， 蔗糖是惰性物质， 能隔绝空气， BSA 和蔗糖可作为支持框架结构的物质。 封闭液的作用使固相载体的其他部位不能 结合蛋白质， 从而保证实验数据的准确性。
本发明在制备方法中缓冲液配方的改进与原发明相比降低非特异性吸附， 减少背景 信号值； 并增加了蛋白芯片的稳定性， 提高了检测的灵敏度。 附图说明
图 1为本发明改进缓冲液配方的制备方法制备的蛋白芯片信号图象。
图 2为原发明制备方法制备的蛋白芯片信号图象。 具体实施方式：
在本发明中， 所用固体材料 (例如 BSA、 蔗糖、 酪氨酸和 NaN3)的用量用重量百分比 表示， 所用溶液 (例如 Tween 20、 胎牛血清和 Proc l in液体）的用量用体积百分比表 示。
实施例一、 用改良后的缓冲液 （包括封闭液、 包被液） 配方制备肿瘤检测蛋白芯片 与以原有缓冲液配方制备的蛋白芯片背景和信号值比较
1、 用改进缓冲液配方的蛋白芯片制备方法制备蛋白芯片 E:
1. 1将 AFP、 CBA、 PSA、 f ree-PSA, CA125 , CA15-3六种肿瘤标志物的对应抗体 (一抗） 以一定浓度溶于包被液中， 然后用芯片自动点样系统将这些蛋白质点 制在固相载体上， 点样密度 48 点 /cm2, 点样量 0. 5ng/点， 每种一抗对应点 4 个平行点；
1. 2 4 °C放置过夜；
1. 3 用封闭液将蛋白芯片封闭 1小时；
1. 4 干燥处理 2小时， 储存于 4 °C备用。
其中用到的包被液配方为 pH为 7. 8的 PB (KH2P04-Na2HP04)。
其中用到的封闭液配方为: 3%BSA, 5%蔗糖, 0. 5。/。。NaN3的 PB (KH2P04- Na2HP04)。 2、 用原蛋白芯片制备方法制备蛋白芯片 F
其中用到的包被液配方为 pH为 9. 6的 CBS (NaHC03-Na2C03)。
其中用到的封闭液配方为：含 0. 2%Tween20 , 0. 1%酪氨酸， 5%BSA , 4%蔗糖， 0. 5°/oproc l in的 TBS。
将上述蛋白芯片 ^ F与配套的反应液、 洗液、 和已知浓度的血清样品反应， （剩 余的血清样品留待下一次实验使用）。 并用生物芯片检测仪进行拍摄和数据分析。信 号值如表 1所示：
表 1
Figure imgf000005_0001
将上述芯片 4 °C贮存放置， 6个月后与取上述实验中同一批号地反应液、 洗液、 和上 述的剩余血清样品反应， 信号值如表 2所示：
表 2
Figure imgf000005_0002
与蛋白芯片 F相比， 按本发明的缓冲液配方制备的蛋白芯片 E其稳定性明显增加。
另外， 由生物芯片拍摄出的蛋白芯片 E信号图象见图 1 , 蛋白芯片 F信号图象 见图 2。 由两图可见， 图 1的背景明显低于图 2。 实施例二、 用改良后的标准品缓冲液配方制备标准品与原有标准品稳定性比较
1. 以如下配方配制标准品缓冲液： 60%胎牛血清 +各种高浓度纯化抗原 +0. 05%。NaN3,另加入 60 μ 1 AFP, CEA, NSE高浓度纯化抗原制成标准品溶液 A;
2. 以 如 下 配 方 配 制 标 准 品 缓 冲 液 ： ΡΗλ 8 0. 05Μ PB ( H2P04- Na2HP04) +15°/oBSA+2. 5%蔗糖 +0. 05%。NaN3,另加入 60 μ 1 AFP, CEA, NSE高浓度 纯化抗原制成标准品溶液 B;
3. 按原有配方配制标准品缓冲液， 另加入 60 μ 1 AFP，CEA, NSE 高浓度纯化抗 原制成标准品溶液 C。 '
注： 溶液 A、 B、 C中的 AFP、 CEA、 NSE终浓度是一致的。
将以上三种标准品溶液冻干， 分别是标准品 a、 b和 c , 用化学发光自动分析仪 进行标定， 标定的浓度值结果如表 3:
表 3
Figure imgf000006_0001
将&、 b、 c三种标准品贮存于 4 °C , 放置 3个月后， 用同一台化学发光自动分析 仪进行复标， 标定的浓度值结果如表 4 :
表 4
Figure imgf000006_0002
与原有缓冲体系制备的标准品 c相比， 标准品 a、 b的浓度更为稳定， 可保存更 长的时间不丧失活性。

Claims

杈利要求
1. 一种多指标并行检测的蛋白芯片检测系统， 由用于多指标并行检测的蛋白芯 片、 以一定浓度配比制成的， 并带有发光标记的多蛋白混合液即反应液、 一系列巳 知浓度且浓度递增的被测蛋白的混合液即标准品、 和洗涤液组成， 其特征在于其中 所述的标准品溶液配方为： 40%- 60%胎牛血清 +各种高浓度纯化抗原 +0. 02-0. l%oNaN3 或 PH7. 0-7. 8 0. 05M PB (KH2PO4-Na2HPO4) +2-30¾BSA+l. 5-2. 5%蔗糖 +0. 02-0. l%oNaN3+ 各种高浓度纯化抗原。
2. 一种如权利要求 1所述的蛋白芯片检测系统的制备方法，其中蛋白芯片制备 包括下列步骤：
1 )确定多种不同的被测物蛋白 （简称目标蛋白 A )和 A特异性的抗原、 抗体或 受体等 （简称 B ), 如体内疾病标志性蛋白和能与之特异结合的多种蛋白；
2 )将上述所述 B即不同的蛋白探针以一定浓度溶于包被液中，然后用芯片自动 点样系统将这些蛋白质点制在固相载体上， 点样密度 25-200 点 /cm2, 点样量 0. 1- 10ng/点；
3 ) 4°C放置过夜；
4 )用封闭液将蛋白芯片封闭；
5 ) 干燥处理， 储存于 4 °C ;
其特征在于其中所述的包被液配方为： pH为 7. 0-8. 0的 PB (KH2P04-Na2HP04)。
3. 一种如杈利要求 1所述的蛋白芯片检测系统的制备方法，其特征在于其中所 述的封闭液配方为： 含 1- 9%BSA， 1 - 9%蔗糖, 0. 01-l%。NaN3的 PB (KH2P04 - Na2HP04)。
PCT/CN2003/000208 2002-10-24 2003-03-24 Systeme de detection de cristaux de proteine pouvant detecter simultanement plusieurs cibles WO2004038413A1 (fr)

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US10/532,837 US20060008793A1 (en) 2002-10-24 2003-03-24 Protein chips detecting system which can simultaneously detect multi target
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102830233A (zh) * 2011-06-13 2012-12-19 上海铭源数康生物芯片有限公司 一种基于硝酸纤维素膜的elisa反应方法
CN107478631A (zh) * 2017-09-19 2017-12-15 南京工业大学 一种同时检测多种肿瘤标志物的3d折叠纸基微流体荧光检测装置
CN116162538A (zh) * 2022-12-16 2023-05-26 中国科学院苏州生物医学工程技术研究所 一种同时检测蛋白和rna的微流控芯片及试剂盒

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010014685A (ja) * 2008-07-04 2010-01-21 Bio Matrix Research Inc タンパク質安定化溶液
CN101609096B (zh) * 2009-07-21 2012-11-07 上海师范大学 肺癌标志物检测免疫层析试纸的制备方法
CN102141569A (zh) * 2010-12-15 2011-08-03 中国疾病预防控制中心寄生虫病预防控制所 一种用于联合诊断三种吸虫病的蛋白芯片及其制备方法
CN108279302B (zh) * 2017-07-11 2020-05-26 深圳市伯劳特生物制品有限公司 一种用于酶联免疫试剂盒的组合物以及幽门螺旋杆菌抗体谱检测试剂盒及其制备方法
CN108414743B (zh) * 2018-03-07 2020-05-26 深圳市伯劳特生物制品有限公司 一种用于酶联免疫试剂盒的组合物以及肿瘤标志物检测试剂盒及其制备方法
CN113030479B (zh) * 2019-12-25 2023-09-12 洛阳中科生物芯片技术有限公司 一种蛋白质芯片的点样溶液

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001014425A1 (en) * 1999-08-19 2001-03-01 Diachip Limited Multipurpose diagnostic systems using protein chips
CN1338634A (zh) * 2001-09-29 2002-03-06 上海晶泰生物技术有限公司 恶性肿瘤早期诊断的蛋白芯片
CN1363840A (zh) * 2001-01-04 2002-08-14 上海数康生物科技有限公司 蛋白芯片及其制备方法和使用方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001014425A1 (en) * 1999-08-19 2001-03-01 Diachip Limited Multipurpose diagnostic systems using protein chips
CN1363840A (zh) * 2001-01-04 2002-08-14 上海数康生物科技有限公司 蛋白芯片及其制备方法和使用方法
CN1338634A (zh) * 2001-09-29 2002-03-06 上海晶泰生物技术有限公司 恶性肿瘤早期诊断的蛋白芯片

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ARENKOV, PAVEL ET AL.: "Protein microchips:Use for Immunoassay and Enzymatic Reactions", ANALYTICAL BIOCHEMISTRY, no. 278, 2000, pages 123 - 131, XP002234264 *
See also references of EP1560023A4 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102830233A (zh) * 2011-06-13 2012-12-19 上海铭源数康生物芯片有限公司 一种基于硝酸纤维素膜的elisa反应方法
CN102830233B (zh) * 2011-06-13 2015-08-26 上海铭源数康生物芯片有限公司 一种基于硝酸纤维素膜的elisa反应方法
CN107478631A (zh) * 2017-09-19 2017-12-15 南京工业大学 一种同时检测多种肿瘤标志物的3d折叠纸基微流体荧光检测装置
CN107478631B (zh) * 2017-09-19 2019-10-11 南京工业大学 一种同时检测多种肿瘤标志物的3d折叠纸基微流体荧光检测装置
CN116162538A (zh) * 2022-12-16 2023-05-26 中国科学院苏州生物医学工程技术研究所 一种同时检测蛋白和rna的微流控芯片及试剂盒
CN116162538B (zh) * 2022-12-16 2024-01-26 中国科学院苏州生物医学工程技术研究所 一种同时检测蛋白和rna的微流控芯片及试剂盒

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