CN101923088A - Gold nanorod immunoprobe, and preparation method and application thereof - Google Patents
Gold nanorod immunoprobe, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a gold nanorod immunoprobe applied to various mediums, and a preparation method and application thereof. The gold nanorod immunoprobe is a product obtained by performing the preparation of gold nano seeds, the growth and concentration of a gold nanorod, antibody modification and sealing. The immunoprobe is subjected to specific recognition in various mediums, such as buffer solution, serum, blood plasma or urine, saliva and the like, and is finally qualitatively and quantitatively analyzed by reading the offset of longitudinal plasma absorption peaks of the gold nanorod. When used for detecting corresponding antigens, the immunoprobe has the advantages of simple operation, high detection sensitivity, high specificity, fewer required instruments and devices, and the like. Moreover, as nano composites are well adapted to external mediums by the special modification mode of the immunoprobe, the immunoprobe also has an expanded application range and is possibly popularized to clinical application.
Description
Technical field
The invention belongs to the nano immune technical field, particularly relate to a kind of gold nanorod immunoprobe that is applicable to multiple medium and preparation method thereof and application.
Background technology
Immunology detection is being taken on crucial role in diagnosis science of today, such as to diagnosis of the detecting in early days of tumour, infectious diseases and immunity disease etc.The ultimate principle of immunology detection is the non-covalent association reaction under multiple power effect between antigen and the antibody, and initial immune response is only carried out between antigen-antibody.Along with the development of related discipline, especially the development that burns the wind to docimasiology of material science has brought new opportunity.Pass through correlated response, the either party of antigen and antibody all can be connected corresponding novel nano label, by immunoreactive generation to the change of character such as the magnetics of label, optics, electricity as monitoring means, play immunoreactive amplification, promote the level of sensitivity of immune detection.As quantum dot, gold nano-material, carbon nano-tube, magnetic nano-particle etc., gold nanorods is exactly wherein comparatively enliven a kind of.
Compare with color of spherical gold, the bar-shaped material of gold nano has some significant advantages: as the absorption of gold nanorods and scattering cross-section than the high order of magnitude of spherical gold grain with volume; The peak position at vertical plasma resonance peak of gold nanorods can simply be implemented in the adjustable continuously of visible and near infrared spectral range by changing its length-diameter ratio; Gold nanorods is higher than spherical gold grain far away to the susceptibility of surrounding environment variations in refractive index.Among the present invention, be exactly the theoretical foundation that its this optical characteristics is studied as nano-biosensing.Generally speaking, the peak of material surface plasma resonance feature moves Δ λ and the refraction index of the sensitivity factor of material, surperficial adsorbed molecule and dissolve medium, relevant (the Nature Materials of factors such as net thickness of adsorbed layer, 2008, vol (7), 442-453).When immune response took place around the gold nanorods, the combination of antigen-antibody will inevitably cause above-mentioned all multifactor variations, thereby changed its sensitive relatively vertical plasma absorption peak position.So far, have many research groups to carry out correlative study in the world, but the most number average among them adopt the method coupling antibody and the gold nanorods of chemical modification, operates comparatively loaded down with trivial details.And previous experiments proves that also the gold nanorods after the chemical modification is poor to the tolerance of media environment, high ionic strength still can cause assembling (the biotechnology communication, 2009, vol 20,680-682).This point is compared with the golden nanometer particle that is widely used (collaurum) and to be had certain gap.
On the other hand, make a general survey of current research, rare research with the application extension of gold nanorod immunoprobe to and clinical relevant medium, as in serum, urine, the saliva (Anal.Chem., 2007,79 (14), 5278-5283).In fact, the research that novel nano-material is applied to the biology sensor aspect is vast as the open sea, but because characteristics such as the surface of nano material itself and interfacial effect, small-size effect, quantum size effect, the development and the application of these sensors are restricted, are in the laboratory development stage for a long time.Specifically, the detection medium of its associated biomolecule sensor seldom can break away from the hotbed of " buffer system ", gets rid of in the actual detected medium interference such as compositions such as albumen, glucose, inorganic ions, enzymes, and enters the clinical practice stage.
Summary of the invention
Be that one of purpose of the present invention provides a kind of gold nanorod immunoprobe that is applicable to multiple medium at the limitation of original nano biological sensor to the detection medium.
The product that this gold nanorod immunoprobe is the preparation of preparation, gold nanorods through the gold nano seed and concentrate, obtain after antibody modification and the sealing.
Second purpose of the present invention provides a kind of preparation method of gold nanorod immunoprobe.
Preparation method provided by the present invention can may further comprise the steps:
1) preparation gold nano seed: under the situation that surfactant hexadecyl trimethyl ammonium bromide (CTAB) exists, the positive trivalent gold in the gold chloride is reduced to the gold nano seed by chemical reduction method;
2) preparation of gold nanorods and concentrate: the gold nano seed of step 1) preparation is joined in the growth solution of gold nanorods and prepare gold nanorods, the CTAB that centrifugal removal is excessive, and gold nanorods concentrated;
3) antibody modification: the gold nanorods after will concentrating joins in the antibody-solutions, makes between gold nanorods and the antibody to react by electrostatic interaction, obtains the gold nanorods-antibody complex with antibody coupling;
4) seal: gold nanorods-antibody complex is washed as sealer with irrelevant protein solution,, obtain the gold nanorod immunoprobe of energy specific recognition corresponding antigens again through centrifuging, concentrated, purifying.
In the preparation method of above-mentioned gold nanorod immunoprobe, the preparation method of described step 1) gold nano seed is specially: at first, the chlorauric acid solution of 0.25mL 0.01mol/L is joined in the CTAB solution of 7.5mL 0.1mol/L, behind the mixing again with 0.6mL 0.01mol/L prepared fresh, place the sodium borohydride solution of ice-water bath to join above-mentioned solution, be inverted mixing back and forth 2 minutes, and obtained the bright brown yellow solution of gold nano seed.
The gold nano seed of described step 1) preparation should be statically placed in 25 ℃ of water-baths, and preparation back 2～5h can use.
Described step 2) method of gold nanorods growth is specially: the chlorauric acid solution of 4mL 10mmol/L, the liquor argenti nitratis ophthalmicus of 0.6mL10mmol/L and the ascorbic acid solution of 0.64mL 0.01mol/L are joined in the CTAB solution of 95mL0.01mol/L successively, then to the 0.1mL gold nano seed solution that wherein adds the step 1) preparation, mixing obtains gold nanorods after still aging; Described aging method is meant and is statically placed in 25 ℃ of water-baths, and the time is 3h at least; The number of times of the excessive CTAB of described centrifugal removal is 2 times, and rotating speed is 8000 rev/mins, and the time is 10min, after described simmer down to is for the second time centrifugal is original 1/20th with the volume-diminished of gold nanorods colloidal solution.
The method of described step 3) antibody modification is specially: 0.2mL gold nanorods concentrate is joined in the antibody-solutions that 1.0mL concentration is 0.1mg/mL, leave standstill 30min behind the mixing, obtain gold nanorods-antibody complex.
Irrelevant protein solution in the described step 4) is the 1%wt bovine serum albumin solution; Sealing is meant to use through the 1%wt of 8000 rev/mins of centrifugal 15min purifying bovine serum albumin solution gold nanorods-antibody complex is washed 3 times, concrete grammar is: the 1%wt bovine serum albumin solution is added in gold nanorods-antibody complex, leave standstill behind the 15min in 4000 rev/mins of centrifugings, supernatant discarded, taking off a layer thing, to add water resuspended, use 1%wt bovine serum albumin solution repeated washing 2 times again, at last gold nanorods-the antibody complex that obtains is concentrated, and be resuspended in the Tris damping fluid of pH7.40.01mol/L, 800 rev/mins of centrifugal 3min purifying, the aggregation that removal may exist, get supernatant and be resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
Another object of the present invention provides the detection method of modified antibodies corresponding antigens on a kind of and the gold nanorod immunoprobe.
The detection method that provides of the present invention, be that above-mentioned gold nanorod immunoprobe is joined in the detected medium, hatch, again with medium and gold nanorod immunoprobe centrifuging, read the skew of the vertical plasma absorption peak that detects back gold nanorod immunoprobe and blank or negative control the result is carried out interpretation (calculated difference Δ λ (nm)), to carry out qualitative and quantitative analysis.
Detection method of the present invention is applicable to that the corresponding antigens of clinical samples such as damping fluid (water or PBS, TBS, Tris etc.), serum, blood plasma, urine, saliva multiple medium commonly used detects.
Described incubation temperature is preferably 37 ℃, and incubation time is preferably 1h.
The described difference DELTA λ that calculates (nm) then thinks systematic error or experiment noise if in ± 3nm, is not judged to be positive findings.
Aqueous media (being in water or the buffer solution), can reaching the high range of linearity of repeatedly parallel laboratory test acquisition reliability by standardization and carry out quantitative test preceding step.
The invention provides a kind of gold nanorod immunoprobe that is applicable to multiple medium and its production and application.With the gold nanorod immunoprobe of the inventive method preparation since between itself and antibody " the full encirclement " formula modification mode of electrostatic interaction (supports of Zata current potential characterization result) and sealer than complete closed, can around gold nanorods, form the shell of the flexibility that constitutes by albumen, make it all various other components in the medium insensitive (reducing the susceptibility of gold nanorods) surrounding medium, thereby realize that this immunological probe detects the highly sensitive and high specific of corresponding antigens, enlarges the range of application of this immunological probe; The specific recognition of this immunological probe can be carried out in multiple medium such as buffer solution, serum, blood plasma or urine, saliva etc., finally carries out qualitative and quantitative analysis by the skew of reading the vertical plasma absorption peak of gold nanorods.The detection of carrying out corresponding antigens with this immunological probe has advantages such as simple to operate, that detection sensitivity is high, specificity good, required instrument and equipment is few, and, because the modification mode that it is special, make nano-complex to external world medium have good adaptability, enlarged the range of application of this immunological probe, the possibility of oriented clinical expansion.
Below in conjunction with specific embodiment the present invention is described in further details.
Description of drawings
The transmission electron microscope photo of the regular gold nanorods that Fig. 1 prepares for the present invention;
" gold nanorods-human IgG " immunological probe that Fig. 2 prepares for the present invention detects the anti-human IgG spectrum peak curve of deviation of variable concentrations in the Tris damping fluid;
" gold nanorods-hepatitis B surface antibody " immunological probe that Fig. 3 prepares for the present invention detects the spectrum peak skew amount effect relation curve of hepatitis B surface antigen standard substance in the Tris damping fluid;
" gold nanorods-hepatitis B surface antibody " immunological probe that Fig. 4 prepares for the present invention detects the spectrum peak skew of hepatitis B surface antigen yin and yang attribute serum, (a) is full figure, (b) is the crest partial enlarged drawing;
The plasma abosrption spectrogram of " gold nanorods-hepatitis B surface antibody " immunological probe in saliva and urine medium that Fig. 5 prepares for the present invention.
Embodiment
Present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.Any the association area experienced person, can utilize other similar antigen-antibody or reaction conditions to realize immunological probe described in the invention according to the principle of this patent.These do not break away from the key concept that the present invention describes.Therefore, these modifications or different application are all within coverage of the present invention.
Method therefor is conventional method if no special instructions among the following embodiment.
The preparation of embodiment 1, " gold nanorods-HIgG " immunological probe with in the Tris damping fluid, detect anti-HIgG
One, the preparation of " gold nanorods-HIgG " immunological probe
With method preparation " gold nanorods-HIgG " immunological probe of the present invention, concrete grammar may further comprise the steps:
(1) preparation gold nano seed: at first, the chlorauric acid solution of 0.25mL 0.01mol/L is joined in the CTAB solution of 7.5mL 0.1mol/L, behind the mixing again with 0.6mL 0.01mol/L prepared fresh, place the sodium borohydride solution of ice-water bath to join above-mentioned solution, be inverted mixing back and forth 2 minutes, and obtained the bright brown yellow solution of gold nano seed.The gold nano seed is statically placed in 25 ℃ of water-baths, uses behind the 3h.
(2) preparation of gold nanorods and concentrated: the chlorauric acid solution of 4mL 10mmol/L, the liquor argenti nitratis ophthalmicus of 0.6mL 10mmol/L and the ascorbic acid solution of 0.64mL 0.01mol/L are joined in the CTAB solution of 95mL 0.01mol/L successively, then to the gold nano seed solution that wherein adds 0.1mL step (1) preparation, mixing, be statically placed in 25 ℃ of water-baths, use behind the ageing 3h.Use preceding 8000 rev/mins centrifugal twice, each 10min to be removing excessive CTAB, and after for the second time centrifugal is original 1/20th with the volume-diminished of gold nanorods colloidal solution.The transmission electron microscope photo of gold nanorods as shown in Figure 1.
(3) antibody modification: the gold nanorods concentrate that 0.2mL step (2) is obtained joins in the HIgG aqueous solution of 1.0mL 0.1mg/mL, leaves standstill 30min behind the mixing, obtains gold nanorods-antibody complex.
(4) sealing: the configuration 10%wt bovine serum albumin solution and in 8000 rev/mins of centrifugal 15min, getting supernatant adds in gold nanorods-antibody complex that step (3) obtains, the final concentration that makes the bovine serum albumin(BSA) sealer is 1%wt, leave standstill behind the 15min in 4000 rev/mins of centrifugings, supernatant discarded, taking off a layer thing, to add water resuspended, again with the bovine serum albumin solution repeated washing of 1%wt 2 times, at last gold nanorods-the antibody complex that obtains is concentrated, and be resuspended in the Tris damping fluid of pH7.4 0.01mol/L, 800 rev/mins of centrifugal 3min purifying, the aggregation that removal may exist, get supernatant and be resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
Two, in the Tris damping fluid, detect anti-HIgG with " gold nanorods-HIgG " immunological probe
(1) the anti-HIgG solution of the Tris damping fluid configuration series concentration of usefulness pH7.4 0.01mol/L.
(2) the anti-HIgG solution of variable concentrations to be measured is added respectively in the gold nanorod immunoprobe liquid of step 1 preparation, make that vertical plasma absorption peak O.D. value of gold nanorods is about 1.0 in the detection architecture.Hatch centrifuging behind the 1h in 37 ℃ of water-baths, remove unreacted albumen, again that lower floor's thing is resuspended in the Tris damping fluid.Adopt ultraviolet-visible spectrophotometer to scan the surface plasma absorption spectrum of the 400-1000nm of each sample.Its Δ λ (nm) is calculated in the difference of gold nanorod immunoprobe liquid before observing the vertical plasma absorption peak of gold nanorods position and detecting, and carries out qualitative or quantitative test.If the offset λ (nm) of the sample that calculates can think systematic error or experiment noise in ± 3nm, be not judged to be positive findings.
The typical case detects spectrogram as shown in Figure 2, promptly can detect the anti-HIgG of nanogram level in the Tris damping fluid.
Preparation of embodiment 2, " gold nanorods-hepatitis B surface antibody " immunological probe and the detection in the Tris damping fluid
One, the preparation of " gold nanorods-hepatitis B surface antibody " immunological probe
With method preparation " gold nanorods-hepatitis B surface antibody " immunological probe of the present invention, concrete grammar may further comprise the steps:
(1) preparation gold nano seed: identical with embodiment 1.
(2) preparation of gold nanorods and concentrated: identical with embodiment 1.
(3) antibody modification: 0.2mL gold nanorods concentrate is joined in the monoclonal hepatitis B surface antibody aqueous solution of 1.0mL 0.1mg/mL, leave standstill 30min behind the mixing, obtain gold nanorods-antibody complex.
(4) sealing: identical with embodiment 1.
Two, in the Tris damping fluid, detect hepatitis B surface antigen with " gold nanorods-hepatitis B surface antibody " immunological probe
(1) with the solution of hepatitis B surface antigen (HBsAg) standard substance of the Tris damping fluid of pH7.4 0.01mol/L configuration series concentration, and the bovine serum albumin solution of 250ng/mL or 500ng/mL.
(2) sample to be tested, dummy (water), control sample (bovine serum albumin solution described in (1)) are added respectively in " gold nanorods-hepatitis B surface antibody " immunological probe liquid of step 1 preparation, make that vertical plasma absorption peak O.D. value of gold nanorods is about 0.5 in the detection architecture.Establish three parallel laboratory tests for every group.Hatch centrifuging behind the 1h in 37 ℃ of water-baths, remove unreacted albumen, again that lower floor's thing is resuspended in the Tris damping fluid.Adopt ultraviolet-visible spectrophotometer to scan the surface plasma absorption spectrum of the 400-1000nm of each sample.Observe the difference of the peak position mean value of gold nanorods vertical plasma absorption peak position and dummy or control sample, calculate its Δ λ (nm), carry out qualitative and quantitative analysis.Usually, dummy can coincide preferably with the peak position of control sample, and this can be used as Success in Experiment whether important criterion.If the offset λ (nm) of the sample that calculates can think systematic error or experiment noise in ± 3nm, be not judged to be positive findings.
Reach repeatedly parallel laboratory test by standardization and obtained amount effect relation curve as shown in Figure 3 preceding step.As shown in the figure, be limited to 0.01IU/mL under the detection of this immunological probe to the HBsAg standard substance, promptly reach the picomole magnitude, compare conventional enzyme-linked immunoassay method detection sensitivity and improve two orders of magnitude.
Embodiment 3: the preparation of " gold nanorods-hepatitis B surface antibody " immunological probe reaches the interpretation to hepatitis B surface antigen positive and negative serum
One, the preparation of " gold nanorods-hepatitis B surface antibody " immunological probe
With method preparation " gold nanorods-hepatitis B surface antibody " immunological probe of the present invention, concrete grammar may further comprise the steps:
(1) preparation gold nano seed: identical with embodiment 1.
(2) preparation of gold nanorods and concentrated: identical with embodiment 1.
(3) antibody modification: identical with embodiment 2.
(4) sealing: identical with embodiment 1.
Two, with of the interpretation of " gold nanorods-hepatitis B surface antibody " immunological probe to hepatitis B surface antigen positive and negative serum
(1) from hepatitis B surface antigen detection kit (available from Yingke Xinchuang (Ximen) Sci. ﹠ Tech. Co., Ltd.), chooses the positive and negative control serum as sample.
(2) yin and yang attribute serum sample to be measured, dummy (water) and control sample (bovine serum albumin solutions in embodiment 2 step 2 (1)) are added respectively in the gold nanorod immunoprobe liquid, make that vertical plasma absorption peak O.D. value of gold nanorods is about 0.5 in the detection architecture.Establish three parallel laboratory tests for every group.Hatch centrifuging behind the 1h in 37 ℃ of water-baths, remove unreacted reactant, again that lower floor's thing is resuspended in the Tris damping fluid.Adopt ultraviolet-visible spectrophotometer to scan the surface plasma absorption spectrum of the 400-1000nm of each sample.Observe the difference of the peak position mean value of gold nanorods vertical plasma absorption peak position and dummy or control sample, calculate its Δ λ (nm), carry out qualitative analysis.Usually, the average value difference in peak position of negative serum, dummy and control sample is in ± 3nm, and the peak position of positive serum and differing of they are at least more than the 8nm.
That Fig. 4 detects the typical plasma abosrption spectrogram (a) of hepatitis B surface antigen positive and negative serum and spectrum peak deviation post for " gold nanorods-hepatitis B surface antibody " immunological probe after the normalization is partial enlarged drawing (b), wherein 1 is dummy, 2 are the control sample, and 3 and 4 are respectively positive serum and negative serum.
Embodiment 4: preparation of " gold nanorods-hepatitis B surface antibody " immunological probe and the surface plasma absorption spectra property in saliva, urine medium thereof
One, the preparation of " gold nanorods-hepatitis B surface antibody " immunological probe
With method preparation " gold nanorods-hepatitis B surface antibody " immunological probe of the present invention, concrete grammar may further comprise the steps:
(1) preparation gold nano seed: identical with embodiment 1.
(2) preparation of gold nanorods and concentrated: identical with embodiment 1.
(3) antibody modification: identical with embodiment 2.
(4) sealing: identical with embodiment 1.
Two, the surface plasma absorption spectra property of " gold nanorods-hepatitis B surface antibody " immunological probe in saliva, urine medium
The gold nanorod immunoprobe liquid for preparing in the step 1 is scattered in respectively in 10% and 40% saliva (saliva) or urine (urine) medium, obtains surface plasma abosrption spectrogram as shown in Figure 5.As seen from Figure 5, in above-mentioned two media, gold nanorod immunoprobe is the energy stable existence all, and particularly its vertical plasma resonance absorption peak position can not be subjected to the influence of medium color, viscosity etc.Can infer that thus the gold nanorod immunoprobe for preparing with the inventive method can be used to detect corresponding antigens/antibody in clinical typical media such as saliva, urine.
Claims (12)
1. the product that gold nanorod immunoprobe is the preparation of preparation, gold nanorods through the gold nano seed and concentrate, obtain after antibody modification and the sealing.
2. the preparation method of a gold nanorod immunoprobe may further comprise the steps:
1) preparation gold nano seed: in the presence of surfactant hexadecyl trimethyl ammonium bromide (CTAB), the positive trivalent gold in the chlorauric acid solution is reduced to the gold nano seed by chemical reduction method;
2) preparation of gold nanorods and concentrate: the gold nano seed of step 1) preparation is joined in the growth solution of gold nanorods and prepare gold nanorods, the CTAB that centrifugal removal is excessive, and gold nanorods concentrated;
3) antibody modification: the gold nanorods after will concentrating joins in the antibody-solutions, makes between gold nanorods and the antibody to react by electrostatic interaction, obtains the gold nanorods-antibody complex with antibody coupling;
4) seal: gold nanorods-antibody complex is washed as sealer with irrelevant protein solution,, obtain the gold nanorod immunoprobe of energy specific recognition corresponding antigens again through centrifuging, concentrated, purifying.
3. preparation method according to claim 2, it is characterized in that: the preparation method of described step 1) gold nano seed is specially: at first, the chlorauric acid solution of 0.25mL 0.01mol/L is joined in the CTAB solution of 7.5mL 0.1mol/L, behind the mixing again with 0.6mL 0.01mol/L prepared fresh, place the sodium borohydride solution of ice-water bath to join above-mentioned solution, be inverted mixing back and forth 2 minutes, and obtained the bright brown yellow solution of gold nano seed.
4. preparation method according to claim 3 is characterized in that: the gold nano seed of described step 1) preparation should be statically placed in 25 ℃ of water-baths, and preparation back 2～5h can use.
5. preparation method according to claim 2, it is characterized in that: described step 2) method of gold nanorods growth is specially: the chlorauric acid solution of 4mL 10mmol/L, the liquor argenti nitratis ophthalmicus of 0.6mL 10mmol/L and the ascorbic acid solution of 0.64mL0.01mol/L are joined in the CTAB solution of 95mL 0.01mol/L successively, then to the 0.1mL gold nano seed solution that wherein adds the step 1) preparation, mixing obtains gold nanorods after still aging; Described aging method is meant and is statically placed in 25 ℃ of water-baths, and the time is 3h at least; The number of times of the excessive CTAB of described centrifugal removal is 2 times, and rotating speed is 8000 rev/mins, and the time is 10min, after described simmer down to is for the second time centrifugal is original 1/20th with the volume-diminished of gold nanorods colloidal solution.
6. preparation method according to claim 2, it is characterized in that: the method for described step 3) antibody modification is specially: 0.2mL gold nanorods concentrate is joined in the antibody-solutions of 0.1mg/mL of 1.0mL concentration, leave standstill 30min behind the mixing, obtain gold nanorods-antibody complex.
7. preparation method according to claim 2 is characterized in that: the irrelevant protein solution in the described step 4) is the 1%wt bovine serum albumin solution; Sealing is meant to use through the 1%wt of 8000 rev/mins of centrifugal 15min purifying bovine serum albumin solution gold nanorods-antibody complex is washed 3 times, concrete grammar is: the 1%wt bovine serum albumin solution is added in gold nanorods-antibody complex, leave standstill behind the 15min in 4000 rev/mins of centrifugings, supernatant discarded, taking off a layer thing, to add water resuspended, use 1%wt bovine serum albumin solution repeated washing 2 times again, at last gold nanorods-the antibody complex that obtains is concentrated, and be resuspended in the Tris damping fluid of pH7.4 0.01mol/L, 800 rev/mins of centrifugal 3min purifying, the aggregation that removal may exist, get supernatant and be resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
One kind with gold nanorod immunoprobe on the detection method of modified antibodies corresponding antigens, be that the described gold nanorod immunoprobe of claim 1 is joined in the detected medium, hatch, again with medium and gold nanorod immunoprobe centrifuging, read the skew of the vertical plasma absorption peak that detects back gold nanorod immunoprobe and blank or negative control the result is carried out interpretation (calculated difference Δ λ (nm)), to carry out qualitative and quantitative analysis.
9. detection method according to claim 8 is characterized in that: described detection method is applicable to that the corresponding antigens of damping fluid (water or PBS, TBS, Tris), serum, blood plasma, urine, saliva medium detects.
10. detection method according to claim 8 is characterized in that: described incubation temperature is 37 ℃, and incubation time is 1h.
11. detection method according to claim 8 is characterized in that: the described difference DELTA λ that calculates (nm) then thinks systematic error or experiment noise if in ± 3nm, is not judged to be positive findings.
12. detection method according to claim 8 is characterized in that:, can reach the high range of linearity of repeatedly parallel laboratory test acquisition reliability by standardization and carry out quantitative test to preceding step at aqueous media.
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| CN103884693A (en) * | 2012-12-20 | 2014-06-25 | 江南大学 | Preparation method for monodispersed and low-biotoxicity gold nanorods, and use for detection of allergen |
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| CN105738342A (en) * | 2016-02-26 | 2016-07-06 | 中国人民解放军军事医学科学院军事兽医研究所 | SERS method for in-situ-synthesizing nano-silver by serving aptamer as support |
| CN105738342B (en) * | 2016-02-26 | 2018-09-25 | 中国人民解放军军事医学科学院军事兽医研究所 | It is a kind of using aptamers as the SERS methods of holder fabricated in situ nano silver |
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| CN109837083A (en) * | 2017-11-24 | 2019-06-04 | 中国农业大学 | Difunctional fluorescence nano flower of a kind of hybrid and the preparation method and application thereof |
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| CN112415079A (en) * | 2019-08-22 | 2021-02-26 | 四川大学 | Double-parameter self-verification homogeneous immunoassay method for single-particle inductively coupled plasma mass spectrometry |
| CN112415079B (en) * | 2019-08-22 | 2021-10-22 | 四川大学 | Double-parameter self-verification homogeneous immunoassay method for single-particle inductively coupled plasma mass spectrometry |
| CN111624347A (en) * | 2020-05-11 | 2020-09-04 | 大理大学 | Application of gold nanorods in detecting serum circulating antigen by specific antibody of liver fluke |
| CN112535886A (en) * | 2020-11-12 | 2021-03-23 | 杭州苏铂科技有限公司 | Method for removing CTAB in gold nanorod solution |
| CN112535886B (en) * | 2020-11-12 | 2022-07-12 | 杭州苏铂科技有限公司 | Method for removing CTAB in gold nanorod solution |
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