CN101493457A - Method for detecting detection objects marked by single-wall carbon nanotube by Raman spectrometer - Google Patents
Method for detecting detection objects marked by single-wall carbon nanotube by Raman spectrometer Download PDFInfo
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- CN101493457A CN101493457A CNA2009100782972A CN200910078297A CN101493457A CN 101493457 A CN101493457 A CN 101493457A CN A2009100782972 A CNA2009100782972 A CN A2009100782972A CN 200910078297 A CN200910078297 A CN 200910078297A CN 101493457 A CN101493457 A CN 101493457A
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- carbon nanotube
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- walled carbon
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Abstract
The invention relates to a method which uses a Raman spectrometer for detecting a detected object that is marked by a single-walled carbon nanotube; firstly the antigens, antibodies or complementary chain DNA biological molecules which can be specifically recognized by an object to be detected are fixed on a single-walled carbon nanotube for obtaining a labeled probe; then the obtained labeled probe is placed in nutrient solution; later, the solution of the object to be detected is dropped on a substrate for forming a detection carrier; next, the nutrient solution which contains at least one labeled probe is dropped on the detection carrier for conducting immune complex reaction; and then deionized water is used for full rinse; after the labeled probes which do not conduct immune complex reaction are removed and then dried by nitrogen, an assembly to be detected is obtained; then 200-1000nm laser is used for exciting the assembly to be detected and the Raman spectra is observed on a Raman spectrometer for observing whether the neighboring places of 150-300cm<-1>, 1350cm<-1>. and 1600cm<-1> wave number have surface-enhanced Raman reflection signals; the invention relates to a novel immune marking method which takes a single-walled carbon nanotube as the marker, and uses the antigens, antibodies or single-chain DNA biological molecules for specific reaction.
Description
Technical field
The invention belongs to field of immunodetection, be particularly related to a kind of antigen, antibody or single stranded DNA of using Single Walled Carbon Nanotube (SWNT) mark under Raman spectrum (SERS) instrument, detected the method that the surface of detecting antigen, antibody or single stranded DNA biomolecule in the thing strengthens specific reaction.
Background technology
Along with the develop rapidly of laboratory medicine, new method, the new technology of immune labeled mensuration constantly occur.Difference according to label can be divided into: immune analysis methods such as isotope labeling, enzyme labeling, golden mark.
Raman spectrum is a characterization of molecules fingerprint vibrational spectrum, and the molecule of different structure has the Raman peaks of different characteristic vibration.The Raman signal of general molecule is very weak, and (the scattering interface is about 10
-30Cm
-2), application surface strengthens Raman technology and can obtain bigger by 10 than conventional Raman spectrum
6Reinforcing coefficient doubly.
Since finding Single Walled Carbon Nanotube in 1993, its particular structure, physics and chemical characteristic are with a wide range of applications it, are one of focuses of current nano materials research.Demonstrate special advantages at Raman spectrum aspect the structural characterization of Single Walled Carbon Nanotube, Raman peaks peak position that it is unique and intensity can provide the information of aspects such as structure and purity, in the research of carbon nano-tube and derivant material thereof, Raman spectrum has been brought into play very important effect, and it can provide molecule abundant vibration information.
Summary of the invention
The technical problem to be solved in the present invention is exactly, provide a kind of antigen, antibody or single stranded DNA of Single Walled Carbon Nanotube mark used under Raman spectrum (SERS) instrument, detect the method that the surface of detecting antigen, antibody or single stranded DNA biomolecule in the thing strengthens specific reaction, it has improved the sensitivity of immune detection.
In order to finish the application's goal of the invention, the present invention by the following technical solutions:
Of the present invention a kind of with the method for Raman spectrometer detection, wherein through the detection thing of Single Walled Carbon Nanotube mark:
(a) will be fixed on the Single Walled Carbon Nanotube with antigen, antibody or the complementary chain dna biomolecule that thing to be detected is discerned mutually, obtain label probe;
(b) the above-mentioned label probe that obtains is put into nutrient solution;
(c) with the drips of solution of thing to be detected on substrate, form and detect carrier, the above-mentioned nutrient solution that will contain at least one label probe again drips and is detecting on the carrier, carry out immune recombination reaction, with the abundant drip washing of deionized water, remove the label probe that does not carry out immune recombination reaction, then afterwards, dry up with nitrogen, make assembly to be detected;
(d) with the above-mentioned assembly to be detected of laser excitation of 200-1000nm, on Raman spectrometer, observe Raman spectrum, observe at 150-300cm
-1, 1350cm
-1And 1600cm
-1Whether has the surface-enhanced Raman reflected signal near the wave number, if above-mentioned surface-enhanced Raman reflected signal is arranged, prove that then thing to be detected has the biomolecule with antibody, antigen or the single stranded DNA of antigen, antibody or the complementary chain dna biomolecule specific recognition of label probe, if there is not above-mentioned surface-enhanced Raman reflected signal, prove that then thing to be detected does not have the biomolecule of the antibody, antigen or the single stranded DNA that combine with antigen, antibody or the DNA complementary strand biomolecule of label probe.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the antigen of thing specific recognition described and to be detected, antibody or complementary chain dna biomolecule are respectively the biomolecule with thing antibody to be detected, antigen or the corresponding antigen of single stranded DNA biomolecule, antibody or complementary chain dna.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the time of described immune recombination reaction is 0.5-5 hour.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the antigen that thing described and to be detected combines, antibody or complementary chain dna biomolecule are fixed on the Single Walled Carbon Nanotube by covalent bond or non-covalent bond.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the antigen that thing described and to be detected is discerned mutually, antibody or complementary chain dna biomolecule are fixed on the Single Walled Carbon Nanotube by amido link on the Single Walled Carbon Nanotube or ester bond covalent bond.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the antigen of thing specific recognition described and to be detected, antibody or complementary chain dna biomolecule are by being fixed on the Single Walled Carbon Nanotube with the mutual noncovalent interaction of Single Walled Carbon Nanotube.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: the solution of the thing that detects is to contain to make the nutrient solution that detects the thing survival.
Of the present invention a kind of with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, wherein: described nutrient solution is the nutrient solution that makes the biomolecule survival of antigen, antibody, complementary chain dna or single stranded DNA.
The present invention is with the Single Walled Carbon Nanotube thing that serves as a mark, with a kind of novel immune labeled method of antigen, antibody or single stranded DNA biomolecule specific reaction.Because the much detected molecule such as the Raman signal of antigen, antibody or single stranded DNA biomolecule are very weak, itself can not be detected by Surface enhanced raman spectroscopy, can introduce the thing that serves as a mark of the Single Walled Carbon Nanotube with the vibration of unique raman signatures, utilize antigen, antibody or single stranded DNA biomolecule specific immune response, formation has the detection thing of haling graceful signal, thereby reaches the purpose of detection.
Description of drawings
Fig. 1 is the synoptic diagram with the to be detected assembly manufacturing process of Raman spectrometer detection in the method for the detection thing of Single Walled Carbon Nanotube mark of the present invention.
Be elaborated with reference to accompanying drawing below in conjunction with embodiment, deep understanding arranged with purpose, feature and advantage to the inventive method
Embodiment
As shown in Figure 1, of the present invention with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, may further comprise the steps:
(a) will with the antigen of thing specific recognition to be detected, antibody or complementary chain dna biomolecule are fixed on the Single Walled Carbon Nanotube 1, obtain label probe, the antigen of discerning mutually with thing to be detected, antibody or complementary chain dna biomolecule are respectively and thing antibody to be detected, antigen or the corresponding antigen of single stranded DNA biomolecule, the biomolecule of antibody or complementary chain dna, the antigen of discerning mutually with thing to be detected, antibody or complementary chain dna biomolecule are that amido link or ester bond covalent bond are fixed on the Single Walled Carbon Nanotube by covalent bond, perhaps by being fixed on the Single Walled Carbon Nanotube with the Single Walled Carbon Nanotube non-covalent bond;
(b) the above-mentioned label probe that obtains is put into nutrient solution; This nutrient solution is the nutrient solution that the biomolecule of antigen, antibody or complementary chain dna can be survived
(c) with the drips of solution of thing 3 to be detected on substrate 2, form and detect carrier, the above-mentioned nutrient solution that will contain at least one label probe again drips detecting on the carrier, carries out immune recombination reaction 0.5-5 hour, and the inspection in reaction time is surveyed the concrete condition of thing and decided, afterwards with the abundant drip washing of ionized water, remove the label probe that does not carry out immune recombination reaction, then, dry up with nitrogen, make assembly to be detected, the solution of thing 3 to be detected is to contain to make the nutrient solution that detects the thing survival;
(d) with the above-mentioned assembly to be detected of laser excitation of 200-1000nm, on Raman spectrometer, observe Raman spectrum, observe at 150-300cm
-1, 1350cm
-1And 1600cm
-1Whether have the surface-enhanced Raman reflected signal near the wave number, this signal is the spike as the mountain.If above-mentioned surface-enhanced Raman reflected signal is arranged, prove that then thing to be detected has the biomolecule with antibody, antigen or the single stranded DNA of antigen, antibody or the complementary chain dna biomolecule specific recognition of label probe, if there is not above-mentioned surface-enhanced Raman reflected signal, prove that then thing to be detected does not have the biomolecule with antibody, antigen or the single stranded DNA of antigen, antibody or the DNA complementary strand biomolecule specific recognition of label probe.
More than describing is explanation of the invention, is not that institute of the present invention restricted portion is referring to claim to the qualification of invention, and under the situation of spirit of the present invention, the present invention can do any type of modification.
Claims (8)
1. one kind is detected method through the detection thing of Single Walled Carbon Nanotube mark with Raman spectrometer, it is characterized in that:
(a) can be fixed on the Single Walled Carbon Nanotube (1) with antigen, antibody or the complementary chain dna biomolecule of thing specific recognition to be detected, obtain label probe;
(b) the above-mentioned label probe that obtains is put into nutrient solution;
(c) with the drips of solution of thing to be detected (3) on substrate (2), form and detect carrier, the above-mentioned nutrient solution that will contain at least one label probe again drips and is detecting on the carrier, carry out immune recombination reaction, with the abundant drip washing of ionized water, remove the label probe that does not carry out immune recombination reaction, then afterwards, dry up with nitrogen, make assembly to be detected;
(d) with the above-mentioned assembly to be detected of laser excitation of 200-1000nm, on Raman spectrometer, observe Raman spectrum, observe at 150-300cm
-1, 1350cm
-1And 1600cm
-1Whether has the surface-enhanced Raman reflected signal near the wave number, if above-mentioned surface-enhanced Raman reflected signal is arranged, prove that then thing to be detected has the biomolecule of antibody, antigen or the single stranded DNA discerned mutually with antigen, antibody or the complementary chain dna biomolecule of label probe, if there is not above-mentioned surface-enhanced Raman reflected signal, prove that then thing to be detected does not have the biomolecule of antibody, antigen or the single stranded DNA discerned mutually with antigen, antibody or the DNA complementary strand biomolecule of label probe.
2. as claimed in claim 1 with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, it is characterized in that: the antigen that thing described and to be detected is discerned mutually, antibody or complementary chain dna biomolecule are respectively the biomolecule with thing antibody to be detected, antigen or the corresponding antigen of single stranded DNA biomolecule, antibody or complementary chain dna.
3. as claimed in claim 2 with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, it is characterized in that: the time of described immune recombination reaction is 0.5-5 hour.
4. as claimed in claim 3 with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, it is characterized in that: the antigen that thing described and to be detected is discerned mutually, antibody or complementary chain dna biomolecule are fixed on the Single Walled Carbon Nanotube (1) by covalent bond or non-covalent bond method of modifying.
5. as claimed in claim 4 with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, it is characterized in that: the antigen that thing described and to be detected is discerned mutually, antibody or complementary chain dna biomolecule are fixed on the Single Walled Carbon Nanotube (1) by acid amides on the Single Walled Carbon Nanotube (1) or ester group covalent bond.
6. as claimed in claim 4 with the method for Raman spectrometer detection through the detection thing of Single Walled Carbon Nanotube mark, it is characterized in that: the antigen that thing described and to be detected is discerned mutually, antibody or complementary chain dna biomolecule are by being fixed on the Single Walled Carbon Nanotube (1) with the mutual noncovalent interaction of Single Walled Carbon Nanotube (1).
7. as claimed in claim 1ly detect method through the detection thing of Single Walled Carbon Nanotube mark with Raman spectrometer, it is characterized in that: the solution of institute's thing that detects (3) is to contain the nutrient solution that the detection thing is survived.
8. describedly detect method through the detection thing of Single Walled Carbon Nanotube mark with Raman spectrometer as claim 7 or 1, it is characterized in that: described nutrient solution is the nutrient solution that makes the biomolecule survival of antigen, antibody, complementary chain dna or single stranded DNA.
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| CNA2009100782972A CN101493457A (en) | 2009-02-24 | 2009-02-24 | Method for detecting detection objects marked by single-wall carbon nanotube by Raman spectrometer |
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| CNA2009100782972A CN101493457A (en) | 2009-02-24 | 2009-02-24 | Method for detecting detection objects marked by single-wall carbon nanotube by Raman spectrometer |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102115026A (en) * | 2009-12-31 | 2011-07-06 | 清华大学 | One-dimensional nano-structure, preparation method thereof and method for marking by using one-dimensional nano-structure |
| CN102023150B (en) * | 2009-09-15 | 2012-10-10 | 清华大学 | Raman scattering substrate and detection system with same |
| US8384893B2 (en) | 2009-09-15 | 2013-02-26 | Tsinghua University | Raman detecting system and detection method for using the same |
| CN105628672A (en) * | 2015-12-17 | 2016-06-01 | 东南大学 | Method for quickly detecting exosomes through SERS signal |
| WO2017177131A1 (en) * | 2016-04-08 | 2017-10-12 | Memorial Sloan Kettering Cancer Center | Sensors for nucleic acid biomarkers |
| CN111537492A (en) * | 2020-04-30 | 2020-08-14 | 东南大学 | Preparation method of uniform high-sensitivity surface-enhanced Raman spectrum probe, probe prepared by same and application of probe |
-
2009
- 2009-02-24 CN CNA2009100782972A patent/CN101493457A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| ANDREW I. MINETT ET AL.: "Decoration of carbon nanotubes with biological entities for electronic device applications", 《NANOSCIENCE AND NANOTECHNOLOGY》 * |
| CHENGFAN CAO ET AL.: "Optical detection of DNA hybridization using absorption spectra of single-walled carbon nanotubes", 《MATERIALS CHEMISTRY AND PHYSICS》 * |
| 任冬酶 等: "单壁碳纳米管的侧壁共价修饰", 《检验检疫科学》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102023150B (en) * | 2009-09-15 | 2012-10-10 | 清华大学 | Raman scattering substrate and detection system with same |
| US8384893B2 (en) | 2009-09-15 | 2013-02-26 | Tsinghua University | Raman detecting system and detection method for using the same |
| US8804116B2 (en) | 2009-09-15 | 2014-08-12 | Tsinghua University | Surface-enhanced Raman scattering substrate and Raman detecting system having the same |
| CN102115026A (en) * | 2009-12-31 | 2011-07-06 | 清华大学 | One-dimensional nano-structure, preparation method thereof and method for marking by using one-dimensional nano-structure |
| CN105628672A (en) * | 2015-12-17 | 2016-06-01 | 东南大学 | Method for quickly detecting exosomes through SERS signal |
| CN105628672B (en) * | 2015-12-17 | 2019-04-09 | 东南大学 | A method of quickly detecting excretion body by SERS signal |
| WO2017177131A1 (en) * | 2016-04-08 | 2017-10-12 | Memorial Sloan Kettering Cancer Center | Sensors for nucleic acid biomarkers |
| CN111537492A (en) * | 2020-04-30 | 2020-08-14 | 东南大学 | Preparation method of uniform high-sensitivity surface-enhanced Raman spectrum probe, probe prepared by same and application of probe |
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Application publication date: 20090729 |