CN101923088B - 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 medium and preparation method thereof and application.
Background technology
Immunology detection is being taken on very important role in diagnosis science of today, such as diagnosis of the early detection to tumour, infectious diseases and immunity disease etc.The ultimate principle of immunology detection is the non-covalent association reaction under the many kinds of force effect between antigen and 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 as docimasiology of material science has brought new opportunity.Pass through correlated response, antigen is connected the either party and all can be connected corresponding novel nano label with antibody, by immunoreactive generation to the change of the 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 realize as seen reaching the adjustable continuously of near infrared spectral range by changing its length-diameter ratio; Gold nanorods to the susceptibility of surrounding environment variations in refractive index far away higher than spherical gold grain.In the present invention, be exactly with the theoretical foundation of its this optical characteristics as nano-biosensing research.Generally, the peak of material surface plasma resonance feature moves Δ λ and the sensitivity factor of material, adsorb on the surface refraction index of molecule and dissolve medium, relevant (the Nature Materials of the factors such as net thickness of adsorbed layer, 2008, vol (7), 442-453).When occurring around gold nanorods, the combination of antigen-antibody will inevitably cause the variation of above-mentioned factors when immune response, thereby changes its relatively sensitive vertical plasma absorption peak position.So far, have in the world many research groups to carry out correlative study, but the great majority in them all adopt method coupling antibody and the gold nanorods of chemical modification, operate comparatively loaded down with trivial details.And previous experiments proves that also the gold nanorods after chemical modification is poor to the tolerance of media environment, and high ionic strength still can cause gathering, and (the biotechnology communication, 2009, vol 20,680-682).This point, compare and have certain gap with the golden nanometer particle that is widely used (collaurum).
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, 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 due to characteristics such as the surface and interface effect of nano material itself, small-size effect, quantum size effects, the development and application of these sensors is restricted, is in for a long time the laboratory development stage.Specifically, the detection medium of its associated biomolecule sensor seldom can break away from the hotbed of " buffer system ", gets rid of the actual interference that detects compositions such as albumen, glucose, inorganic ions, enzyme in medium, and enters the clinical practice stage.
Summary of the invention
Be that one of purpose of the present invention is to provide a kind of gold nanorod immunoprobe that is applicable to medium for the limitation of original nano biological sensor to the detection medium.
This gold nanorod immunoprobe is the product that obtains after the preparation of preparation, gold nanorods through the gold nano seed and concentrated, antibody modification and sealing.
Second purpose of the present invention is to provide a kind of preparation method of gold nanorod immunoprobe.
Preparation method provided by the present invention can comprise the following steps:
1) preparation gold nano seed:, in the situation that surfactant hexadecyl trimethyl ammonium bromide (CTAB) exists, by chemical reduction method, the positive trivalent gold in gold chloride is reduced to the gold nano seed;
2) preparation of gold nanorods and concentrated: with step 1) the gold nano seed of preparation joins in the growth solution of gold nanorods and prepares gold nanorods, the CTAB that centrifugal removal is excessive, and gold nanorods is concentrated;
3) antibody modification: the gold nanorods after concentrating joins in antibody-solutions, makes between gold nanorods and antibody and reacts 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, then, through centrifuging, concentrated, purifying, obtain the gold nanorod immunoprobe of energy specific recognition corresponding antigens.
In the preparation method of above-mentioned gold nanorod immunoprobe, described step 1) preparation method of 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, again the fresh preparation of 0.6mL 0.01mol/L, the sodium borohydride solution that is placed in ice-water bath are joined above-mentioned solution after mixing, be inverted and mix 2 minutes back and forth, obtain the bright brown yellow solution of gold nano seed.
Described step 1) the gold nano seed of preparation should be statically placed in 25 ℃ of water-baths, and after preparation, 2～5h can use.
Described step 2) method of gold nanorods growth is specially: the ascorbic acid solution of the liquor argenti nitratis ophthalmicus of the chlorauric acid solution of 4mL 10mmol/L, 0.6mL10mmol/L and 0.64mL 0.01mol/L is joined successively in the CTAB solution of 95mL0.01mol/L, then add wherein step 1) preparation 0.1mL gold nano seed solution, mix, obtain gold nanorods after still aging; Described aging method refers to be 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, with the volume-diminished of gold nanorods colloidal solution, is original 1/20th after described simmer down to is centrifugal for the second time.
Described step 3) method of antibody modification is specially: 0.2mL gold nanorods concentrate is joined in the antibody-solutions that 1.0mL concentration is 0.1mg/mL, mix rear standing 30min, obtain gold nanorods-antibody complex.
described step 4) the irrelevant protein solution in is the 1%wt bovine serum albumin solution, sealing refers to use through the 1%wt of 8000 rev/mins of centrifugal 15min purifying bovine serum albumin solution to gold nanorods-antibody complex washing 3 times, concrete grammar is: the 1%wt bovine serum albumin solution is added in gold nanorods-antibody complex, after standing 15min in 4000 rev/mins of centrifugings, supernatant discarded, taking off a layer thing, to add water resuspended, use again 1%wt bovine serum albumin solution repeated washing 2 times, gold nanorods-the antibody complex that will obtain finally 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, getting supernatant is resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
Another object of the present invention is to provide the detection method of modified antibodies corresponding antigens on a kind of and gold nanorod immunoprobe.
The detection method that provides of the present invention, that above-mentioned gold nanorod immunoprobe is joined in detected medium, hatch, again with medium and gold nanorod immunoprobe centrifuging, read the skew of the vertical plasma absorption peak that detects rear gold nanorod immunoprobe and blank or negative control result is carried out interpretation (calculated difference Δ λ (nm)), to carry out qualitative and quantitative analysis.
The corresponding antigens that detection method of the present invention is applicable to the clinical samples such as damping fluid (water or PBS, TBS, Tris etc.), serum, blood plasma, urine, saliva medium commonly used detects.
Described incubation temperature is preferably 37 ℃, and incubation time is preferably 1h.
The described difference DELTA λ that calculates (nm), if in ± 3nm, thinks systematic error or experiment noise, is not judged to be positive findings.
, aqueous media (being in water or buffer solution), can reaching the high range of linearity of repeatedly parallel laboratory test acquisition reliability by the standardization to preceding step and carry out quantitative test.
The invention provides a kind of gold nanorod immunoprobe that is applicable to medium and its preparation method and application.With the gold nanorod immunoprobe of the inventive method preparation because " the full encirclement " formula of electrostatic interaction (supports of Zata current potential characterization result) between itself and antibody is modified sealing more fully of mode and sealer, can form the shell of the flexibility that is formed by albumen around gold nanorods, make it all various other components in medium insensitive (reducing the susceptibility of gold nanorods to 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 buffer solution, serum, blood plasma or the mediums such as urine, saliva, finally by the skew of reading the vertical plasma absorption peak of gold nanorods, carry out qualitative and quantitative analysis.The detection of carrying out corresponding antigens with this immunological probe has the 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
Fig. 1 is the transmission electron microscope photo of the regular gold nanorods prepared of the present invention;
Fig. 2 is " gold nanorods-human IgG " immunological probe that the present invention prepares detects variable concentrations in the Tris damping fluid anti-human IgG spectrum peak curve of deviation;
Fig. 3 is " gold nanorods-hepatitis B surface antibody " immunological probe that the present invention prepares detects the hepatitis B surface antigen standard substance in the Tris damping fluid spectrum peak skew amount effect relation curve;
Fig. 4 is the spectrum peak skew that " gold nanorods-hepatitis B surface antibody " immunological probe of preparing of the present invention detects hepatitis B surface antigen yin and yang attribute serum, (a) is full figure, is (b) the crest partial enlarged drawing;
Fig. 5 is the plasma abosrption spectrogram of " gold nanorods-hepatitis B surface antibody " immunological probe in saliva and urine medium that the present invention prepares.
Embodiment
The present embodiment is implemented under take technical solution of the present invention as prerequisite, 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, according to the principle of this patent, utilize other similar antigen-antibody or reaction conditions to realize immunological probe described in the invention.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.
In following embodiment, method therefor is conventional method if no special instructions.
The preparation of embodiment 1, " gold nanorods-HIgG " immunological probe with detect anti-HIgG in the Tris damping fluid
One, the preparation of " gold nanorods-HIgG " immunological probe
With method preparation " gold nanorods-HIgG " immunological probe of the present invention, concrete grammar comprises the following 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, again the fresh preparation of 0.6mL 0.01mol/L, the sodium borohydride solution that is placed in ice-water bath are joined above-mentioned solution after mixing, be inverted and mix 2 minutes back and forth, obtain the bright brown yellow solution of gold nano seed.The gold nano seed is statically placed in 25 ℃ of water-baths, uses after 3h.
(2) preparation of gold nanorods and concentrated: the ascorbic acid solution of the liquor argenti nitratis ophthalmicus of the chlorauric acid solution of 4mL 10mmol/L, 0.6mL 10mmol/L and 0.64mL 0.01mol/L is joined successively in the CTAB solution of 95mL 0.01mol/L, the gold nano seed solution that then adds wherein 0.1mL step (1) preparation, mix, be statically placed in 25 ℃ of water-baths, use after ageing 3h.Use front 8000 rev/mins centrifugal twice, each 10min to be to remove excessive CTAB, and with the volume-diminished of gold nanorods colloidal solution, is original 1/20th after centrifugal for the second time.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, mixes rear standing 30min, 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, after standing 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, gold nanorods-the antibody complex that will obtain finally 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, getting supernatant is resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
Two, detect anti-HIgG with " gold nanorods-HIgG " immunological probe in the Tris damping fluid
(1) configure the anti-HIgG solution of series concentration with the Tris damping fluid of 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 vertical plasma absorption peak O.D. value of gold nanorods in detection system be about 1.0.Hatch centrifuging after 1h in 37 ℃ of water-baths, remove unreacted albumen, then 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.Before observing the vertical plasma absorption peak of gold nanorods position and detecting, the difference of gold nanorod immunoprobe liquid, calculate its Δ λ (nm), carries out qualitative or quantitative test., if the offset Δ λ of the sample that calculates (nm) in ± 3nm, can think systematic error or experiment noise, be not judged to be positive findings.
The typical case detects spectrogram as shown in Figure 2, namely the anti-HIgG of nanogram level can be detected in the Tris damping fluid.
The 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 comprises the following 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, mix rear standing 30min, obtain gold nanorods-antibody complex.
(4) sealing: identical with embodiment 1.
Two, detect hepatitis B surface antigen with " gold nanorods-hepatitis B surface antibody " immunological probe in the Tris damping fluid
(1) configure the solution of hepatitis B surface antigen (HBsAg) standard substance of series concentration with the Tris damping fluid of pH7.4 0.01mol/L, 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 vertical plasma absorption peak O.D. value of gold nanorods in detection system be about 0.5.Establish three parallel laboratory tests for every group.Hatch centrifuging after 1h in 37 ℃ of water-baths, remove unreacted albumen, then 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 and the peak position meeting quite well of controlling sample, and this can be used as Success in Experiment whether important criterion., if the offset Δ λ of the sample that calculates (nm) in ± 3nm, can think systematic error or experiment noise, be not judged to be positive findings.
Reach repeatedly parallel laboratory test by the standardization to preceding step and obtained amount effect relation curve as shown in Figure 3.As shown in the figure, be limited to 0.01IU/mL under the detection of this immunological probe to the HBsAg standard substance, namely 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 comprises the following 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 the interpretation of " gold nanorods-hepatitis B surface antibody " immunological probe to hepatitis B surface antigen positive and negative serum
(1) choose the positive and negative control serum as sample from hepatitis B surface antigen detection kit (available from Yingke Xinchuang (Ximen) Sci. ﹠ Tech. Co., Ltd.).
(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 gold nanorod immunoprobe liquid, make vertical plasma absorption peak O.D. value of gold nanorods in detection system be about 0.5.Establish three parallel laboratory tests for every group.Hatch centrifuging after 1h in 37 ℃ of water-baths, remove unreacted reactant, then 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, negative serum, dummy and the average value difference in peak position of controlling sample are in ± 3nm, and more than the differing and be at least 8nm of the peak position of positive serum and they.
Fig. 4 be " gold nanorods-hepatitis B surface antibody " immunological probe after normalization detect the typical plasma abosrption spectrogram (a) of hepatitis B surface antigen positive and negative serum and spectrum peak deviation post be partial enlarged drawing (b), wherein 1 is dummy, 2 for controlling sample, and 3 and 4 are respectively positive serum and negative serum.
Embodiment 4: the 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 comprises the following 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 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 subject to the impact of medium color, viscosity etc.Can infer thus, with the gold nanorod immunoprobe of the inventive method preparation, can be used to detect corresponding antigens/antibody in the clinical typical media such as saliva, urine.
Claims (11)
1. the preparation method of a gold nanorod immunoprobe comprises the following steps:
1) preparation gold nano seed: under the existence of surfactant hexadecyl trimethyl ammonium bromide (CTAB), by chemical reduction method, the positive trivalent gold in chlorauric acid solution is reduced to the gold nano seed;
2) preparation of gold nanorods and concentrated: with step 1) the gold nano seed of preparation joins in the growth solution of gold nanorods and prepares gold nanorods, the CTAB that centrifugal removal is excessive, and gold nanorods is concentrated; Concrete operations are: the ascorbic acid solution of the liquor argenti nitratis ophthalmicus of the chlorauric acid solution of 4mL10mmol/L, 0.6mL10mmol/L and 0.64mL0.01mol/L is joined successively in the CTAB solution of 95mL0.01mol/L, then add wherein step 1) preparation 0.1mL gold nano seed solution, mix, obtain gold nanorods after still aging;
3) antibody modification: the 0.2mL gold nanorods after concentrating joins in the antibody-solutions that 1.0mL concentration is 0.1mg/mL, mix rear standing 30min, make between gold nanorods and antibody and react by electrostatic interaction, obtain the gold nanorods-antibody complex with antibody coupling; Described antibody is HIgG or hepatitis B surface antibody;
4) seal: gold nanorods-antibody complex is washed as sealer with irrelevant protein solution, then, through centrifuging, concentrated, purifying, obtain the gold nanorod immunoprobe of energy specific recognition corresponding antigens; Described irrelevant protein solution is the 1wt% bovine serum albumin solution; Sealing refers to use through the 1wt% of 8000 rev/mins of centrifugal 15min purifying bovine serum albumin solution to gold nanorods-antibody complex washing 3 times.
2. preparation method according to claim 1, it is characterized in that: described step 1) the preparation concrete operations of gold nano seed are: at first, the chlorauric acid solution of 0.25mL0.01mol/L is joined in the CTAB solution of 7.5mL0.1mol/L, again the fresh preparation of 0.6mL0.01mol/L, the sodium borohydride solution that is placed in ice-water bath are joined above-mentioned solution after mixing, be inverted and mix 2 minutes back and forth, obtain the bright brown yellow solution of gold nano seed.
3. preparation method according to claim 2 is characterized in that: described step 1) the gold nano seed of preparation should be statically placed in 25 ℃ of water-baths, and after preparation, 2～5h can use.
4. preparation method according to claim 1 is characterized in that: described step 2), ageing refers to be 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, with the volume-diminished of gold nanorods colloidal solution, is original 1/20th after described simmer down to is centrifugal for the second time.
5. preparation method according to claim 1, it is characterized in that: described step 4) concrete operations are: the 1wt% bovine serum albumin solution is added in gold nanorods-antibody complex, after standing 15min in 4000 rev/mins of centrifugings, supernatant discarded, taking off a layer thing, to add water resuspended, use again 1wt% bovine serum albumin solution repeated washing 2 times, gold nanorods-the antibody complex that will obtain finally 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, getting supernatant is resuspended in the Tris damping fluid, obtain gold nanorod immunoprobe liquid.
6. gold nanorod immunoprobe is the product that obtains after the preparation of the preparation of gold nano seed, gold nanorods and concentrated, antibody modification and sealing by the arbitrary described method of claim 1 to 5.
One kind with gold nanorod immunoprobe on the detection method of modified antibodies corresponding antigens, that gold nanorod immunoprobe claimed in claim 6 is joined in detected medium, hatch, again with medium and gold nanorod immunoprobe centrifuging, read the skew of the vertical plasma absorption peak that detects rear gold nanorod immunoprobe and blank or negative control result is carried out interpretation, be calculated difference Δ λ (nm), to carry out qualitative and quantitative analysis.
8. detection method according to claim 7 is characterized in that: the corresponding antigens that described detection method is applicable to damping fluid, serum, blood plasma, urine, saliva medium detects, and described damping fluid is water, PBS, TBS or Tris.
9. detection method according to claim 7, it is characterized in that: described incubation temperature is 37 ℃, incubation time is 1h.
10. detection method according to claim 7 is characterized in that: the described difference DELTA λ that calculates (nm), if in ± 3nm, think systematic error or experiment noise, is not judged to be positive findings.
11. detection method according to claim 7 is characterized in that:, at aqueous media, can reach the high range of linearity of repeatedly parallel laboratory test acquisition reliability by the standardization to preceding step and carry out quantitative test.
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| 人IgG标记金纳米棒并用于抗人IgG抗体的检测;彭剑淳;《生物技术通讯》;20090930;第20卷(第5期);第681页第1.5节，第2.3节 * |
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| 金纳米棒的表面改性及其在生物医学领域的应用;杨玉东等;《化学通报》;20100318(第3期);全文 * |
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| CN101923088A (en) | 2010-12-22 |
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