Method for detecting biomolecule by using Fe3O4@Au nucleocapsid nano-probe

Abstract

A method for detecting a biomolecule by using a Fe3O4@Au nucleocapsid nano-probe relates to a surface-enhanced raman spectroscopy and includes the steps of first preparing a Fe3O4@Au probe; then preparing a gold probe; and finally raman-detecting. The method has the advantages of being capable of extracting, washing and concentrating a target biomolecule from a complex biological sample matrix conveniently by using an applied magnetic field, omitting multifarious operation such as centrifugation, filtering and the like, greatly improving detecting efficiency of the biomolecule, having good chemical stability and biological compatibility, and being capable of being used in detecting of the biomolecule safely. The probe combines the surface-enhanced raman spectroscopy, a detecting system has strong surface-enhanced raman signals, fast high-sensitivity detecting on the biomolecule is achieved by detecting enhanced raman signals of a marked molecule, and detecting sensitivity in an experiment is 10-7M. Compared with a traditional biological method, the method is simple and fast to operate, good in reproductively, and capable of achieving detecting of clinical samples.

Description

technical field

[0001] The invention relates to a method based on surface-enhanced Raman spectroscopy, in particular to a method using Fe 3 o 4 Au core-shell nanoprobes for the rapid detection of biomolecules. Background technique

[0002] The rapid and accurate detection of biomolecules in biological samples has important research significance and practical value for obtaining chemical and biological information in life processes, diagnosing biomedical diseases, and chemical analysis. The detection of biomolecules includes the detection of organs, tissues, cells, and even DNA, proteins and other biomacromolecules. This requires that our detection devices must be gradually miniaturized, and at the same time ensure the high efficiency and sensitivity of detection, which is a great challenge faced by many traditional bioanalysis methods.

[0003] With the rapid development of nanoscience and technology, nanobiological probes composed of nanomaterials have become a new gener...

Examples

Embodiment 1

[0034] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0035] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0036] (2) Preparation of 2.6nm gold nanoparticles: refer to the method [Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313.] method, the specific steps are, take 1ml 1%HAucl 4 The solution was dropped into 90 ml of ultra...

Embodiment 2

[0042] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0043] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0044] (2) Preparation of 2.6nm gold nanoparticles: with reference to the method of literature (Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313), the specific steps are to measure 1ml 1 %HAuCl 4 The solution was droppe...

Embodiment 3

[0050] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0051] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0052] (2) Preparation of 2.6nm gold nanoparticles: refer to the method [Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313.] method, the specific steps are, take 1ml 1%HAuCl 4 The solution was dropped into 90 ml of ultra...

technical field

[0001] The invention relates to a method based on surface-enhanced Raman spectroscopy, in particular to a method using Fe 3 o 4 Au core-shell nanoprobes for the rapid detection of biomolecules. Background technique

[0002] The rapid and accurate detection of biomolecules in biological samples has important research significance and practical value for obtaining chemical and biological information in life processes, diagnosing biomedical diseases, and chemical analysis. The detection of biomolecules includes the detection of organs, tissues, cells, and even DNA, proteins and other biomacromolecules. This requires that our detection devices must be gradually miniaturized, and at the same time ensure the high efficiency and sensitivity of detection, which is a great challenge faced by many traditional bioanalysis methods.

[0003] With the rapid development of nanoscience and technology, nanobiological probes composed of nanomaterials have become a new gener...

Examples

Embodiment 1

[0034] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0035] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0036] (2) Preparation of 2.6nm gold nanoparticles: refer to the method [Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313.] method, the specific steps are, take 1ml 1%HAucl 4 The solution was dropped into 90 ml of ultra...

Embodiment 2

[0042] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0043] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0044] (2) Preparation of 2.6nm gold nanoparticles: with reference to the method of literature (Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313), the specific steps are to measure 1ml 1 %HAuCl 4 The solution was droppe...

Embodiment 3

[0050] 1) Fe 3 o 4 Preparation of Au probe: First, it can be prepared according to the method disclosed in Chinese patent CN101773810A, the specific method is as follows:

[0051] (1) Preparation of iron ferric oxide nanoparticles: weigh 0.26g FeSO 4 ·7H 2 O dissolved in 2.5ml 2M / L KNO 3 Add 2.5ml of 1M / L NaOH solution to the solution, stir, add 20ml of 1-5g / L PEI solution with a molecular weight of (1,800-25,000), put it into a microwave reactor at 90°C, and react for 30-120min. After the reaction, the magnetic separation was washed 5 times with deionized water, dispersed in 25ml water and preserved, and the final gained solution ferric oxide content concentration was 0.012M (mass ratio of FeSO 4 ·7H 2 O: KNO 3 :NaOH=1.87:10:5).

[0052] (2) Preparation of 2.6nm gold nanoparticles: refer to the method [Brown, K.R.; Walter, D.G.; Natan, M.J.Chem.Mater.2000, 12(2), 306-313.] method, the specific steps are, take 1ml 1%HAuCl 4 The solution was dropped into 90 ml of ultra...

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