CN103713139A - Test paper for human heat shock protein (Hsp 90alpha) and preparation method thereof - Google Patents
Test paper for human heat shock protein (Hsp 90alpha) and preparation method thereof Download PDFInfo
- Publication number
- CN103713139A CN103713139A CN201310729382.7A CN201310729382A CN103713139A CN 103713139 A CN103713139 A CN 103713139A CN 201310729382 A CN201310729382 A CN 201310729382A CN 103713139 A CN103713139 A CN 103713139A
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- pad
- test paper
- hsp90
- collaurum
- 90alpha
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- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
-
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- 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
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57415—Specifically defined cancers of breast
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
-
- 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
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57446—Specifically defined cancers of stomach or intestine
Abstract
The invention discloses immunochromatography test paper, and particularly relates to immunochromatography test paper for detecting human heat shock protein (Hsp 90alpha) and a preparation method thereof. The immunochromatography test paper is characterized in that a detection line is arranged on the immunochromatography test paper and is formed by coating an antibody which is a white rat anti-human Hsp 90alpha monoclonal antibody; a quality control line is formed by coating a goat anti-rat IgG polyclonal antibody; a colloidal gold combination pad is prepared by using a rat anti-human Hsp 90alpha monoclonal antibody and colloid gold. The preparation method of the Hsp 90alpha detection immunochromatography test paper comprises the following steps: A. coating a nitrocellulose membrane with an antibody detection line working solution and a quality control line working solution, and drying the obtained nitrocellulose membrane for 8-12 hours at the temperature of 36-38 DEG C, thus obtaining a chromatography membrane; B. preparing the colloidal gold combination pad by using the rat anti-human Hsp 90alpha monoclonal antibody and the colloid gold; and C. pasting the chromatography membrane and the colloidal gold combination pad on a glue plate according to the sequence of a specimen pad, the colloidal gold combination pad, the chromatography membrane and a water absorption pad so as to prepare the immunochromatography test paper for detecting the Hsp 90alpha.
Description
Technical field
The present invention discloses a kind of immune chromatography test paper, is specifically related to immune chromatography test paper of a kind of detection human heat shock protein (Hsp90 α) and preparation method thereof.
Background technology
Immune chromatography test paper is that a kind of chromatographic theory that utilizes grades the target detection molecule in liquid to be detected or one-tenth on test paper, to flow to detection line (T line) and nature controlling line (C line), when detection line generation change color illustrates that testing result is positive, otherwise negative, when nature controlling line generation change color, illustrate that testing result is effective, otherwise invalid.
Immune chromatography test paper has advantage very easily when detecting or diagnosing some disease, so it has fully been applied in the detection or diagnostic procedure of various diseases or symptom.Generally, immune chromatography test paper includes following several structure: sample pad, pad, chromatographic film, adsorptive pads, supporting pad, and described sample pad, collaurum pad, chromatographic film profit adsorptive pads stick on supporting pad successively; In described chromatographic film, be disposed with detection line and nature controlling line; Its principle of work is: drop to be measured is added in sample pad, and liquid to be measured utilizes chromatographic theory to flow to pad, and in liquid to be measured, the collaurum bond of determinand in pad is combined, and jointly flows to successively detection line and the nature controlling line in chromatographic film with it; In liquid to be measured, contain target detection molecule, thus the antibody component that target detection molecule, collaurum bond can comprise in detection line be combined and form immune complex, detection line color changes simultaneously; When liquid to be measured flow to nature controlling line, the antibody component comprising in nature controlling line can be combined with collaurum bond, thereby nature controlling line color changes.
Human heat shock protein 90 α, Hsp90 α, is the important member in heat-shock protein family.External expert's reported first in 1989 gene order of Hsp90 α, confirmed the identity of this albumen.Within 1992, foreign scientist finds, Hsp90 α can be by tumor cell secretion to extracellular, but its Mechanism of Secretion Regulation after this for a long time in and unclear.
Being firmly established at present of this brand-new tumor markers of Hsp90 α, opened the molecular difference of Hsp90 α in extracellular hsp 90 α and cell simultaneously, further illustrate secreting type Hsp90 α and can promote tumor invasion and transfer, and its content and malignancy positive correlation in blood.These discoveries have proved in blood that Hsp90 α is as the good potential quality of tumor markers.Hsp90 α tumor markers has the characteristic of wide spectrum, can be used for the clinical detection of a plurality of knurl kinds such as liver cancer, breast cancer, colorectal cancer, can reflect real-time and accurately result for the treatment of.Hsp90 α, as tumor markers, checks more convenient and quicker, and also everybody reduces cost.Only need get one and bleed, can detect by this kit the content of Hsp90 α in blood plasma.Although some tumor patient does not show related symptoms, but its Hsp90 alpha content may raise, if first detect Hsp90 α level in blood, carry out again testing in depth testing, just may be diagnosed as tumour, this can make patient receive treatment early, thereby everybody improves cure rate or extends life cycle.
In addition, in patient's blood plasma, the height of Hsp90 alpha content has good correspondence with change of illness state, therefore can reflect in real time, more exactly result for the treatment of, for doctor formulates and adjust in time therapeutic scheme, provides reference.
Summary of the invention
In view of this, the invention provides immune chromatography test paper of a kind of Hsp90 of detection α and preparation method thereof, there is the advantages such as sampling is easy, easy and simple to handle, reaction is quick, susceptibility is high, high specificity, applicable Site Detection.
For solving above technical matters, technical scheme of the present invention is to adopt a kind of Hsp90 α to detect immune chromatography test paper, and this immune chromatography test paper includes sample pad, pad, chromatographic film, adsorptive pads and supporting pad; Described sample pad, collaurum pad, chromatographic film, adsorptive pads are connected with supporting pad successively in order; It is characterized in that: in chromatographic film, be disposed with detection zone and Quality Control district, described detection zone is near collaurum pad one end, and Quality Control district is near adsorptive pads one end; In described detection zone, be provided with detection line, described detection line is to form by mouse-anti people Hsp90 alpha monoclonal antibodies is coated; Described Quality Control is provided with nature controlling line in district, and described nature controlling line is to form by sheep anti-mouse igg polyclonal antibody is coated; Described collaurum pad: mouse-anti people Hsp90 alpha monoclonal antibodies and collaurum are prepared into collaurum pad.
In described detection line and nature controlling line, the working fluid concentration of each antibody is mouse-anti people Hsp90 alpha monoclonal antibodies 3.0-2.5mg/mL, sheep anti-mouse igg polyclonal antibody 1.0-2.0mg/mL.
In described detection line and nature controlling line, the working fluid concentration of each antibody is mouse-anti people Hsp90 alpha monoclonal antibodies 2.0mg/mL, sheep anti-mouse igg polyclonal antibody 3.0mg/mL.
The preparation process that described Hsp90 α detects immune chromatography test paper is as follows:
A, each antibody detection line working fluid and nature controlling line working fluid are coated with on nitrocellulose filter, gained nitrocellulose filter at 36-38 ℃ dry 8-12 hour chromatographic film;
B, mouse-anti people Hsp90 alpha monoclonal antibodies and collaurum are prepared into collaurum pad;
C, the chromatographic film of A and B step gained and collaurum pad are sticked on successively and on offset plate, are prepared into described detection Hsp90 alpha immunization chromatographic test paper according to the order of sample pad, collaurum pad, chromatographic film, adsorptive pads.
The invention has the beneficial effects as follows: immune chromatography test paper of a kind of Hsp90 of detection α and preparation method thereof, compare with conventional reagents, have that sampling is easy, easy and simple to handle, reaction fast, high, the high specificity of susceptibility, be applicable to Site Detection reduce testing staff workload etc. advantage.
Accompanying drawing explanation:
Fig. 1 Hsp90 α detection kit test paper structure schematic diagram.
Reference numeral explanation: 1: sample pad; 2: pad; 3: chromatographic film; 4: adsorptive pads; 5: detection line (T); 6: nature controlling line (C); 7: supporting pad.
Embodiment
Embodiment:
A, each antibody detection line working fluid and nature controlling line working fluid are coated with on nitrocellulose filter, gained nitrocellulose filter at 36-38 ℃ dry 8-12 hour chromatographic film;
B, mouse-anti people Hsp90 alpha monoclonal antibodies profit collaurum is prepared into collaurum pad;
C, the chromatographic film of A and B step gained and collaurum pad are sticked on successively and on offset plate, are prepared into described detection Hsp90 alpha immunization chromatographic test paper according to the order of sample pad, collaurum pad, chromatographic film, adsorptive pads.
Claims (4)
1. a Hsp90 α detects immune chromatography test paper and preparation method thereof, and this immune chromatography test paper includes sample pad, pad, chromatographic film, adsorptive pads and supporting pad; Described sample pad, collaurum pad, chromatographic film, adsorptive pads are connected with supporting pad successively in order; It is characterized in that: in chromatographic film, be disposed with detection zone and Quality Control district, described detection zone is near collaurum pad one end, and Quality Control district is near adsorptive pads one end; In described detection zone, be provided with detection line, described detection line is to form by mouse-anti people Hsp90 alpha monoclonal antibodies is coated; Described Quality Control is provided with nature controlling line in district, and described nature controlling line is to form by sheep anti-mouse igg polyclonal antibody is coated; Described collaurum pad: mouse-anti people Hsp90 alpha monoclonal antibodies and collaurum are prepared into collaurum pad.
2. Hsp90 α detects immune chromatography test paper according to claim 1, it is characterized in that: in described detection line and nature controlling line, the working fluid concentration of each antibody is mouse-anti people Hsp90 alpha monoclonal antibodies 0.5-3.0mg/mL, sheep anti-mouse igg polyclonal antibody 2.0-5.0mg/mL.
3. Hsp90 α detects immune chromatography test paper according to claim 2, it is characterized in that: in described detection line and nature controlling line, the working fluid concentration of each antibody is mouse-anti people Hsp90 alpha monoclonal antibodies 2.0mg/mL, sheep anti-mouse igg polyclonal antibody 3.0mg/mL.
4. prepare the preparation method that Hsp90 α described in claim 1 detects immune chromatography test paper, it is characterized in that including following steps:
A, each antibody detection line working fluid and nature controlling line working fluid are coated with on nitrocellulose filter, gained nitrocellulose filter at 36-38 ℃ dry 8-12 hour chromatographic film;
B, mouse-anti people Hsp90 alpha monoclonal antibodies and collaurum are prepared into collaurum pad;
C, the chromatographic film of A and B step gained and collaurum pad are sticked on successively and on offset plate, are prepared into described detection Hsp90 alpha immunization chromatographic test paper according to the order of sample pad, collaurum pad, chromatographic film, adsorptive pads.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310729382.7A CN103713139A (en) | 2013-12-14 | 2013-12-14 | Test paper for human heat shock protein (Hsp 90alpha) and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310729382.7A CN103713139A (en) | 2013-12-14 | 2013-12-14 | Test paper for human heat shock protein (Hsp 90alpha) and preparation method thereof |
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| Publication Number | Publication Date |
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| CN103713139A true CN103713139A (en) | 2014-04-09 |
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| CN201310729382.7A Pending CN103713139A (en) | 2013-12-14 | 2013-12-14 | Test paper for human heat shock protein (Hsp 90alpha) and preparation method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403706A (en) * | 2015-04-30 | 2016-03-16 | 北京丹大生物技术有限公司 | Heat shock protein 90 (HSP90) chemiluminescence detection kit |
| CN106248957A (en) * | 2016-07-15 | 2016-12-21 | 陶少强 | A kind of biochip for detecting HUMAN HEAT SHOCK PROTEINS Hhsp HSP70 |
| CN108896761A (en) * | 2018-06-29 | 2018-11-27 | 广州爱康生物技术有限公司 | The detection card of the luxuriant and rich with fragrance blue shape bacterium of Marni a kind of, detection box and preparation method thereof |
| CN110684105A (en) * | 2019-09-11 | 2020-01-14 | 江苏莱森生物科技研究院有限公司 | anti-HSP 90 monoclonal antibody and kit |
| CN111879933A (en) * | 2020-07-30 | 2020-11-03 | 广州德成生物科技有限公司 | Immunochromatography test paper for detecting novel coronavirus |
| CN116925219A (en) * | 2023-09-19 | 2023-10-24 | 北京索莱宝科技有限公司 | Antibody of small heat shock protein HSPB1, hybridoma cell strain and application thereof |
| CN117347635A (en) * | 2023-10-08 | 2024-01-05 | 烟台普罗吉生物科技发展有限公司 | Heat shock protein 90 alpha chemiluminescence detection kit and preparation method thereof |
| CN117388494A (en) * | 2023-10-08 | 2024-01-12 | 烟台普罗吉生物科技发展有限公司 | Application of tumor marker in tumor diagnosis and tumor treatment monitoring products |
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Patent Citations (1)
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| KR20030092378A (en) * | 2002-05-29 | 2003-12-06 | 박영미 | Method and kit for diagnosis of progress stage of solid cancer |
Non-Patent Citations (2)
| Title |
|---|
| 东星等: "大肠癌中HSP70和HSP90α表达与其生物学行为的关系", 《中国普通外科杂志》, vol. 20, no. 10, 31 October 2011 (2011-10-31), pages 1120 - 1122 * |
| 刘志刚等: "M2-PK和HSP90α在胃癌患者血清中的表达及意义", 《世界华人消化杂志》, vol. 19, no. 25, 8 September 2011 (2011-09-08), pages 2673 - 2678 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403706A (en) * | 2015-04-30 | 2016-03-16 | 北京丹大生物技术有限公司 | Heat shock protein 90 (HSP90) chemiluminescence detection kit |
| CN106248957A (en) * | 2016-07-15 | 2016-12-21 | 陶少强 | A kind of biochip for detecting HUMAN HEAT SHOCK PROTEINS Hhsp HSP70 |
| CN106248957B (en) * | 2016-07-15 | 2018-07-03 | 安徽国科生物科技有限公司 | A kind of biochip for being used to detect human heat shock protein |
| CN108896761A (en) * | 2018-06-29 | 2018-11-27 | 广州爱康生物技术有限公司 | The detection card of the luxuriant and rich with fragrance blue shape bacterium of Marni a kind of, detection box and preparation method thereof |
| CN110684105A (en) * | 2019-09-11 | 2020-01-14 | 江苏莱森生物科技研究院有限公司 | anti-HSP 90 monoclonal antibody and kit |
| CN111879933A (en) * | 2020-07-30 | 2020-11-03 | 广州德成生物科技有限公司 | Immunochromatography test paper for detecting novel coronavirus |
| CN111879933B (en) * | 2020-07-30 | 2023-10-03 | 广州德成生物科技有限公司 | Immunochromatography test paper for detecting novel coronavirus |
| CN116925219A (en) * | 2023-09-19 | 2023-10-24 | 北京索莱宝科技有限公司 | Antibody of small heat shock protein HSPB1, hybridoma cell strain and application thereof |
| CN116925219B (en) * | 2023-09-19 | 2023-12-08 | 北京索莱宝科技有限公司 | Antibody of small heat shock protein HSPB1, hybridoma cell strain and application thereof |
| CN117347635A (en) * | 2023-10-08 | 2024-01-05 | 烟台普罗吉生物科技发展有限公司 | Heat shock protein 90 alpha chemiluminescence detection kit and preparation method thereof |
| CN117388494A (en) * | 2023-10-08 | 2024-01-12 | 烟台普罗吉生物科技发展有限公司 | Application of tumor marker in tumor diagnosis and tumor treatment monitoring products |
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Application publication date: 20140409 |