CN109900897A - A kind of optimization method that the lateral flow biosensor based on golden platinum nano flower tests and analyzes - Google Patents
A kind of optimization method that the lateral flow biosensor based on golden platinum nano flower tests and analyzes Download PDFInfo
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Abstract
The invention belongs to immunoassays and other biological detection technique field, more particularly to the optimization method that a kind of lateral flow biosensor based on golden platinum nano flower tests and analyzes, the optimization including surveying optimization to gold platinum nano flower solution ph used when being used to prepare golden platinum nano flower-antibody coupling matter, to golden platinum nano flower-antibody coupling matter solution usage, p-wire preparation condition in antibody dosage when being used to prepare golden platinum nano flower-antibody coupling matter, sample solution.The present invention is had the advantage that in the present invention compared with prior art by optimizing to detection and analysis process, it can make the lateral flow biosensor based on golden platinum nano flower that there is optimum performance, lateral flow biosensor can be enhanced to low concentration intentional Protein Detection sensitivity compared with prior art by generating；The Protein Detection that can be used in bio-matrix can also further expand to other biological monitoring, such as DNA and microRNA, have biggish promotion potential.
Description
Technical field
The invention belongs to immunoassays and other biological detection technique field, and in particular to a kind of based on golden platinum nano flower
The optimization method that lateral flow biosensor tests and analyzes.
Background technique
Lateral flow immunoassay is a kind of paper base biosensor of real time monitoring, since its is at low cost, operation letter
The advantages that single and be concerned, another major advantage of lateral flow immunoassay is that it can be enterprising in various biological samples
Row, including blood plasma, sweat, saliva, serum, urine and whole blood, in addition, for detecting required sample size than needed for conventional detection
Detect much smaller, therefore, lateral flow immunoassay has been used extensively as a kind of very useful portable analysis tool
In the quick detection of protein；Currently, including gold nano grain, quantum dot, magnetic nanoparticle, up-conversion luminescence nano particle
The label of lateral flow immunoassay all it has been used as with the various nano particles including active Raman nano material, in these materials
In material, gold nano grain is most widely used label, because they have unique optical property and easy surface modification, so
And due to the finite surface area of gold nano grain, the lateral flow immunoassay based on gold nano grain cannot detect physiological liquid
The protein biomarker object of certain low concentrations in body.Other nanoparticle labels relevant to high sensor really may be used
To greatly improve the sensitivity of lateral flow immunoassay, but unfortunately these quantitative approach are all relied on for complicated letter
Number readout equipment, and these nano material labels are difficult to prepare, therefore, urgent need do not have the highly sensitive of disadvantages mentioned above
Quick detection of the novel lateral flow analytical signal strategy for protein target, and many basic research and medicine are examined
Breaking, there have to be very useful.
In order to which low cost detects protein rapidly and sensitively, we propose a kind of side based on golden platinum nano flower (APN) first
To flowing immuno analytical method, during this investigation it turned out, we by growing Pt nanowires on gold nano grain surface being prepared
Golden platinum nano flower be used as lateral flow immunological technique in dual label.There is the dual mark of colorimetric and catalysis in golden platinum nano flower
In the case where label, golden platinum nano flower is trapped on test strip, showing darkened features band, (golden platinum nano flower is as colorimetric
Label), golden platinum nano flower can further be catalyzed the chromogenic substrate of addition and generate color products to enhance the intensity of test strip
(golden platinum nano flower is as nano enzyme label)；It finds during the experiment, different experiment conditions will affect the inspection of target protein
It surveys, therefore, in order to obtain the optimum performance of the lateral flow biosensor (LFB) based on golden platinum nano flower, needs to detection
The condition of analytic process advanced optimizes.
Summary of the invention
The purpose of the present invention is being directed to existing issue, a kind of lateral flow bio-sensing based on golden platinum nano flower is provided
The optimization method that device tests and analyzes.
The present invention is achieved by the following technical solutions: a kind of lateral flow biosensor based on golden platinum nano flower
The optimization method of detection and analysis, including marker is connected with antibody and prepares marker-antibody coupling matter, then with target protein
After mixing is immunoreacted, target protein is detected by lateral flow biosensor, the marker is golden platinum
Nano flower, target protein are rabbit immunoglobulin, and the antibody is goat anti-rabbit igg；Including anti-to golden platinum nano flower-is used to prepare
Gold platinum nano flower solution ph used when body conjugate, to antibody dosage when being used to prepare golden platinum nano flower-antibody coupling matter, right
Golden platinum nano flower-antibody coupling matter solution usage, the p-wire preparation condition to lateral flow biosensor in sample solution
Optimization.
It includes the following contents that a kind of lateral flow biosensor based on golden platinum nano flower, which tests and analyzes process:
(1) golden platinum nano flower-antibody coupling matter is prepared
(1.1) 500 microlitres of mass concentrations of centrifuge washing are 1.0mg/mL under conditions of 9000 revs/min, are discarded after the completion
Gained precipitating is resuspended in 1.0mL water, while adjusting pH value by clear liquid, and it is molten to obtain the golden platinum nano flower that pH value is 7.0-10.0
Liquid；
(1.2) antibody that 15-25 microlitres of mass concentration is 1.0mg/mL is added in golden platinum nano flower solution, and in room temperature condition
Under be stirred to react 2 hours, obtain reaction solution；
(1.3) it is then added after being equivalent to the ethanol amine of its weight 5% and stirs 30 minutes in reaction solution, then add phase
When small in the BSA stirring 1 of reaction solution weight 10%, mixed liquor is obtained；
(1.4) gained mixed liquor is centrifuged 10 minutes under conditions of 8000 revs/min, discards supernatant liquid, precipitating is resuspended in
In washing buffer；
(1.5) step (1.4) are repeated three times, then gained precipitating is resuspended in elution buffer, the elution buffer packet
Sodium phosphate buffer containing 20mM, 5wt% bovine serum albumin(BSA), 10wt% sucrose and 0.25wt% Tween-20, then by gained gold platinum
Nano flower-antibody coupling matter solution stores for future use under conditions of 4 DEG C；
(2) lateral flow biosensor analysis target protein is utilized, the lateral flow biosensor includes sample pad, nitre
Acid cellulose film and water absorption pad, the sample pad, nitrocellulose filter and water absorption pad successively overlap and are assembled in viscous plastic back
On liner plate, each overlap is at least overlapped 2mm or more, is sprayed on goat anti-rabbit igg as p-wire on nitrocellulose filter, spray
Film repeats 3-6 times, is sprayed on nitrocellulose filter using the anti-sheep IgG of donkey as control line；
(2.1) in sample-loading buffer, by 100 microlitres of rabbit immunoglobulin sample solutions containing various concentration and 1-3 microlitres
Golden platinum nano flower-antibody coupling matter solution mixing, obtains mixture；
(2.2) gained mixture drop is mobile towards water absorption pad in sample pad, after ten minutes, add 60 microlitres of loading buffer
Liquid washs lateral flow biosensor；
(2.3) visually rank p-wire and control line in 20 minutes, the image of shooting lateral flow biosensor is for fixed
Property analysis；
(2.4) it using the intensity of portable bar shaped reader record p-wire and control line, the i.e. peak area of curve obtained, is used for
Quantitative analysis.
As further improvement of these options, the washing buffer contains 10mMPBS and 1wt%BSA；The loading
Buffer PBS, 1wt%BSA and 0.1wt%PVP containing 10mM.
As further improvement of these options, the Jin Bona for being used to prepare golden platinum nano flower-antibody coupling matter
Popped rice pH value of solution is 9；The antibody for being used to prepare golden platinum nano flower-antibody coupling matter is equivalent to the dosage of golden platinum nano flower solution and is
25μg/mL；Described to be sprayed on goat anti-rabbit igg as p-wire on nitrocellulose filter, spray film is repeated 5 times；The step (3.1)
The middle rabbit immunoglobulin sample solution by 100 microlitres containing various concentration and 2.5 microlitres of golden platinum nano flower-antibody coupling matters are molten
Liquid mixing, signal-to-noise ratio (S/N) ratio histogram of the LFB by experimental result based on APN, obtaining this is optimal conditions, can
Make the lateral flow biosensor based on golden platinum nano flower that there is optimum performance.
As further improvement of these options, by the sample-loading buffer of step (2.1), in 10wt% human plasma
Rabbit immunoglobulin is added, the rabbit immunoglobulin sample solution of various concentration is prepared；In order to assess exploitation based on APN's
Whether the protein detection that LFB is applied in bio-matrix is feasible, and test is added to the human plasma of the rabbit igg of various concentration, leads to
The percentage of human plasma in change sample solution is crossed to study matrix effect；It was found that there is no base when blood plasma percentage is lower than 10%
Mass effect, this feasibility for being used to that the LFB based on APN to be examined to detect target protein.
Further include step (2.5) as further improvement of these options, contains in the addition of lateral flow biosensor
The colour developing of the colour developing bottom liquid of 3- amino -9- Ethy-Carbazole and hydrogen peroxide, the image for then shooting lateral flow biosensor are used
It is used in qualitative analysis using the intensity of portable bar shaped reader record p-wire and control line, the i.e. peak area of curve obtained
In quantitative analysis.
The present invention has the advantage that compared with prior art by optimizing to detection and analysis process in the present invention, energy
Make the lateral flow biosensor based on golden platinum nano flower that there is optimum performance, in the case where being not added with chromogenic substrate,
Target protein concentration is that there are visible test strips by 0.5ng/mL, after adding chromogenic substrate, is in target protein concentration
There are visible test strips by 0.05ng/mL, and lateral flow biosensor can be enhanced to low concentration mesh compared with prior art by generating
Mark Protein Detection sensitivity；And have verified that the Protein Detection that can be used in bio-matrix, not shadow in the presence of 10% blood plasma
Analysis performance is rung, other biological monitoring, such as DNA and microRNA can be also further expanded to, there is biggish promotion potential.
Detailed description of the invention
Fig. 1 is the detection and analysis process of the lateral flow biosensor based on golden platinum nano flower.
Fig. 2 is experimental condition optimization content.
Fig. 3 is signal-to-noise ratio (S/N) ratio histogram of the LFB based on APN in the presence of target protein and non-targeted albumen.
Fig. 4 be add chromogenic substrate before calibration tape peak area and human plasma in target protein concentration relationship figure, wherein inserting
Figure is the respective alignment curve between the peak area of calibration tape and the logarithm of object rabbit igg concentration；
Fig. 5 is illustration in Fig. 4；
Fig. 6 be add chromogenic substrate before calibration tape peak area and human plasma in target protein concentration relationship figure, wherein illustration be
Respective alignment curve between the peak area of calibration tape and the logarithm of object rabbit igg concentration；
Fig. 7 is illustration in Fig. 6.
Specific embodiment
The following further describes the present invention with reference to the drawings.
When implementing, agents useful for same source is as follows: gold chloride tetrahydrate (HAuCl4.4H2O) have purchased from lark prestige science and technology
Limit company (BeiJing, China)；3- amino -9- Ethy-Carbazole (AEC), hydrogenperoxide steam generator (H2O2), ethanol amine, bovine serum albumin
White (BSA), sucrose, polysorbas20, rabbit igg, phosphate buffer solution (0.1M PBS, pH7.4) is purchased from the raw work bioengineering in Shanghai
Co., Ltd's (Chinese Shanghai)；Goat anti-rabbit igg (GaR IgG, Ab1) and the anti-sheep IgG(Ab2 of donkey) it is limited purchased from the rich biology difficult to understand in Beijing
Company (BeiJing, China).Glass fiber sample pad, polyvinyl chloride (PVC) viscous plastic backer board, cellulose fiber peacekeeping nitric acid are fine
It ties up plain film (Pall 90) and is purchased from Shanghai Jie Ning Bioisystech Co., Ltd (Chinese Shanghai).
Part Experiment instrument and purposes include: that Canon's EOS 1000D camera (Canon, Japan) is raw for shooting lateral flow
The photograph image of object sensor；Use the portable test-strips bought from Shanghai Jinbiao Bio-Tech Co., Ltd.'s (Chinese Shanghai)
Reader (DT2032) collects quantitative data；With the silent winged generation of microplate reader 1510(match, you are scientific and technological) measurement ultra-violet absorption spectrum；It uses
The microgram of JEM-2100F transmission electron microscope (Tokyo) shooting Jenner's popped rices of Hitachi.
The detection and analysis process of lateral flow biosensor as shown in fig. 1 based on golden platinum nano flower, such as Fig. 2 (A)
For the optimization to golden platinum nano flower solution ph used when being used to prepare golden platinum nano flower-antibody coupling matter, if Fig. 2 (B) is pair
Antibody dosage when being used to prepare golden platinum nano flower-antibody coupling matter；If Fig. 2 (C) is that golden platinum nano flower-antibody is even in sample solution
Join the optimization of object solution usage；If Fig. 2 (D) is that p-wire preparation condition surveys optimization；
A kind of optimization method that the lateral flow biosensor based on golden platinum nano flower tests and analyzes, including by marker and resist
Body, which is connected, prepares marker-antibody coupling matter, then mixes with target protein after being immunoreacted, and passes through lateral flow biology
Sensor detects target protein, and the marker is golden platinum nano flower, and target protein is rabbit immunoglobulin, described anti-
Body is goat anti-rabbit igg, including the following contents:
(1) golden platinum nano flower-antibody coupling matter is prepared
(1.1) 500 microlitres of mass concentrations of centrifuge washing are 1.0mg/mL under conditions of 9000 revs/min, are discarded after the completion
Gained precipitating is resuspended in 1.0mL water, while adjusting pH value by clear liquid, obtains the golden platinum nano flower solution that pH value is 9.0；
(1.2) antibody that 25 microlitres of mass concentrations are 1.0mg/mL is added in golden platinum nano flower solution, and at room temperature
It is stirred to react 2 hours, obtains reaction solution；
(1.3) it is then added after being equivalent to the ethanol amine of its weight 5% and stirs 30 minutes in reaction solution, then add phase
When small in the BSA stirring 1 of reaction solution weight 10%, mixed liquor is obtained；
(1.4) gained mixed liquor is centrifuged 10 minutes under conditions of 8000 revs/min, discards supernatant liquid, precipitating is resuspended in
In washing buffer；
Wherein, washing buffer contains 10mMPBS and 1wt%BSA；
(1.5) step (1.4) are repeated three times, then gained precipitating is resuspended in elution buffer, the elution buffer packet
Sodium phosphate buffer containing 20mM, 5wt% bovine serum albumin(BSA), 10wt% sucrose and 0.25wt% Tween-20, then by gained gold platinum
Nano flower-antibody coupling matter solution stores for future use under conditions of 4 DEG C；
(2) lateral flow biosensor analysis target protein is utilized, the lateral flow biosensor includes sample pad, nitre
Acid cellulose film and water absorption pad, the sample pad, nitrocellulose filter and water absorption pad successively overlap and are assembled in viscous plastic back
On liner plate, each overlap is at least overlapped 2mm or more, is sprayed on goat anti-rabbit igg as p-wire on nitrocellulose filter, spray
Film is repeated 5 times, and is sprayed on nitrocellulose filter using the anti-sheep IgG of donkey as control line；
(2.1) in sample-loading buffer, by 100 microlitres of rabbit immunoglobulin sample solutions containing various concentration and 2.5 microlitres
Golden platinum nano flower-antibody coupling matter solution mixing, obtains mixture；
Wherein, PBS, 1wt%BSA and 0.1wt%PVP containing 10mM of sample-loading buffer；
(2.2) gained mixture drop is mobile towards water absorption pad in sample pad, after ten minutes, add 60 microlitres of loading buffer
Liquid washs lateral flow biosensor；
(2.3) visually rank p-wire and control line in 20 minutes, the image of shooting lateral flow biosensor is for fixed
Property analysis；
(2.4) it using the intensity of portable bar shaped reader record p-wire and control line, the i.e. peak area of curve obtained, is used for
Quantitative analysis；
(2.5) it is shown in the colour developing bottom liquid that lateral flow biosensor adds the -9- of amino containing 3- Ethy-Carbazole and hydrogen peroxide
Color, the image for then shooting lateral flow biosensor are used for qualitative analysis, are recorded and tested using portable bar shaped reader
The intensity of line and control line, the i.e. peak area of curve obtained are used for quantitative analysis.
Wherein, the preparation method of the golden platinum nano flower is that the 1wt% chlorauric acid solution of 0.5mL is added to 50mL water
In solution, gold chloride prefabricated solution is obtained, is heated to boiling, and under agitation, by the 1wt% sodium citrate solution of 0.8mL
It rapidly joins；Until there is golden red solution, it is again heated to boiling, the 0.1M ascorbic acid of 1mL is added, adds 1.25
The 1wt% platinum acid chloride solution of mL, after continuous heating 25 minutes to get.
In addition, with rabbit immunoglobulin is added in 10wt% human plasma, preparation is not by the sample-loading buffer of step (3.1)
With the rabbit immunoglobulin sample solution of concentration；In order to assess the egg being applied to the LFB based on APN of exploitation in bio-matrix
White matter detects whether feasible, and test is added to the human plasma of the rabbit igg of various concentration, passes through and changes human plasma in sample solution
Percentage studies matrix effect；It was found that not having matrix effect when blood plasma percentage is lower than 10%, this is based on APN for examining
LFB detection target protein feasibility.
Setting experiment 1
The specificity that rabbit immunoglobulin is detected for confirming the LFB based on APN；
Experimental group 1 and control group are set, and experimental group 1-5 is the rabbit immunoglobulin sample solution that mass concentration is 5ng/mL, right
Alpha-fetoprotein, carcinomebryonic antigen, fibrin ferment, human serum albumins and the junket that mass concentration is 50ng/mL are replaced with respectively according to group 1-5
Albumen obtains significant height in the presence of rabbit immunoglobulin sample solution and responds as shown in Figure 3, and other groups
It obtains insignificant corresponding, illustrate the LFB based on APN to rabbit immunoglobulin detection with high degree of specificity.
Setting experiment 2
It is whether feasible for assessing the protein detection being applied to the LFB based on APN of exploitation in bio-matrix
In test, test is added to the human plasma of the rabbit igg of various concentration, by the percentage for changing human plasma in sample solution
Than studying matrix effect, discovery does not have matrix effect when blood plasma percentage is lower than 10%, this is for examining based on APN's
The feasibility of LFB detection target protein.
Fig. 4 be add chromogenic substrate before calibration tape peak area and human plasma in target protein concentration relationship figure, wherein inserting
Figure i.e. Fig. 5 is the respective alignment curve between the peak area of calibration tape and the logarithm of object rabbit igg concentration；Fig. 6 is that addition is aobvious
Target protein concentration relationship figure in the peak area and human plasma of calibration tape before color substrate, wherein illustration, that is, Fig. 7 is the peak of calibration tape
Respective alignment curve between area and the logarithm of object rabbit igg concentration；
As can be seen that the dynamic range of the LFB based on APN (is 0.5 before addition chromogenic substrate there are rabbit igg
Ng/mL to 20 ng/mL；Add chromogenic substrate after for 0.05 ng/mL to 100.05 ng/mL) in the concentration of 10% blood plasma with
There is no identical when blood plasma；The above results show that the LFB based on APN can be used for detecting target protein in the presence of complex matrices.
Golden platinum nano flower is prepared by Pt nanowires long on gold nano grain, and obtained golden platinum nano flower is made
For the sensitive-type lateral flow immunoassay of double labeling, in conjunction with the unique optical properties of gold nanoparticle and urging for Pt nanowires
Change performance, prepared golden platinum nano flower double labelling lateral flow measurement experiment is able to detect that minimum concentration is 0.05ng/mL
Target protein, this is 20 times lower than traditional lateral flow measurement experiment based on gold nanoparticle；In addition, based on golden platinum nanometer
Colored lateral flow measurement experiment shows excellent anti-interference, and the analysis performance measured in the presence of 10% blood plasma is not
It is impacted.Therefore, golden platinum nano flower shows that exploitation sensitivity lateral flow experiment connects immunoadsorption assay with nano flower
Huge hope；The application of golden platinum nano flower can extend to the label as other biological detection, such as DNA and microRNA.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Claims (10)
1. the optimization method that a kind of lateral flow biosensor based on golden platinum nano flower tests and analyzes, including by marker with
Antibody, which is connected, prepares marker-antibody coupling matter, then mixes with target protein after being immunoreacted, raw by lateral flow
Object sensor detects target protein, which is characterized in that the marker is golden platinum nano flower, and target protein is immune for rabbit
Globulin, the antibody are goat anti-rabbit igg；Including to golden platinum nanometer used when being used to prepare golden platinum nano flower-antibody coupling matter
Flower solution ph, to antibody dosage when being used to prepare golden platinum nano flower-antibody coupling matter, to platinum nano flower-golden in sample solution
Antibody coupling matter solution usage, the optimization to the p-wire preparation condition of lateral flow biosensor.
2. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as described in claim 1
Method, which is characterized in that it includes the following contents that the lateral flow biosensor based on golden platinum nano flower, which tests and analyzes process:
(1) golden platinum nano flower-antibody coupling matter is prepared
(1.1) 500 microlitres of mass concentrations of centrifuge washing are 1.0mg/mL under conditions of 9000 revs/min, are discarded after the completion
Gained precipitating is resuspended in 1.0mL water, while adjusting pH value by clear liquid, and it is molten to obtain the golden platinum nano flower that pH value is 7.0-10.0
Liquid；
(1.2) antibody that 15-25 microlitres of mass concentration is 1.0mg/mL is added in golden platinum nano flower solution, and in room temperature condition
Under be stirred to react 2 hours, obtain reaction solution；
(1.3) it is then added after being equivalent to the ethanol amine of its weight 5% and stirs 30 minutes in reaction solution, then add phase
When small in the BSA stirring 1 of reaction solution weight 10%, mixed liquor is obtained；
(1.4) gained mixed liquor is centrifuged 10 minutes under conditions of 8000 revs/min, discards supernatant liquid, precipitating is resuspended in
In washing buffer；
(1.5) step (1.4) are repeated three times, then gained precipitating is resuspended in elution buffer, the elution buffer packet
Sodium phosphate buffer containing 20mM, 5wt% bovine serum albumin(BSA), 10wt% sucrose and 0.25wt% Tween-20, then by gained gold platinum
Nano flower-antibody coupling matter solution stores for future use under conditions of 4 DEG C；
(2) lateral flow biosensor analysis target protein is utilized, the lateral flow biosensor includes sample pad, nitre
Acid cellulose film and water absorption pad, the sample pad, nitrocellulose filter and water absorption pad successively overlap and are assembled in viscous plastic back
On liner plate, each overlap is at least overlapped 2mm or more, is sprayed on goat anti-rabbit igg as p-wire on nitrocellulose filter, spray
Film repeats 3-6 times, is sprayed on nitrocellulose filter using the anti-sheep IgG of donkey as control line；
(2.1) in sample-loading buffer, by 100 microlitres of rabbit immunoglobulin sample solutions containing various concentration and 1-3 microlitres
Golden platinum nano flower-antibody coupling matter solution mixing, obtains mixture；
(2.2) gained mixture drop is mobile towards water absorption pad in sample pad, after ten minutes, add 60 microlitres of loading buffer
Liquid washs lateral flow biosensor；
(2.3) visually rank p-wire and control line in 20 minutes, the image of shooting lateral flow biosensor is for fixed
Property analysis；
(2.4) it using the intensity of portable bar shaped reader record p-wire and control line, the i.e. peak area of curve obtained, is used for
Quantitative analysis.
3. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that the washing buffer contains 10mMPBS and 1wt%BSA；The sample-loading buffer PBS, 1wt% containing 10mM
BSA and 0.1wt%PVP.
4. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that the golden platinum nano flower pH value of solution for being used to prepare golden platinum nano flower-antibody coupling matter is 9.
5. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that the antibody for being used to prepare golden platinum nano flower-antibody coupling matter is equivalent to the dosage of golden platinum nano flower solution and is
25μg/mL。
6. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that described to be sprayed on goat anti-rabbit igg as p-wire on nitrocellulose filter, spray film is repeated 5 times.
7. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that by 100 microlitres of rabbit immunoglobulin sample solutions and 2.5 containing various concentration in the step (2.1)
Microlitre golden platinum nano flower-antibody coupling matter solution mixing.
8. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as claimed in claim 2
Method, which is characterized in that in sample-loading buffer, add rabbit immunoglobulin in 10wt% human plasma, prepare the rabbit of various concentration
Immunoglobulin sample solution.
9. a kind of optimization that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as described in claim 1 or 8
Method, which is characterized in that further include step (2.5), add-the 9- of amino containing 3- Ethy-Carbazole in lateral flow biosensor
It develops the color with the colour developing bottom liquid of hydrogen peroxide, the image for then shooting lateral flow biosensor is used for qualitative analysis, using just
The intensity of formula bar shaped reader record p-wire and control line, the i.e. peak area of curve obtained are taken, quantitative analysis is used for.
10. a kind of optimization side that the lateral flow biosensor based on golden platinum nano flower tests and analyzes as described in claim 1
Method, which is characterized in that the preparation method of the gold platinum nano flower is that the 1wt% chlorauric acid solution of 0.5mL is added to 50mL water
In solution, gold chloride prefabricated solution is obtained, is heated to boiling, and under agitation, by the 1wt% sodium citrate solution of 0.8mL
It rapidly joins；Until there is golden red solution, it is again heated to boiling, the 0.1M ascorbic acid of 1mL is added, adds 1.25
The 1wt% platinum acid chloride solution of mL, after continuous heating 25 minutes to get.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106885902A (en) * | 2017-03-03 | 2017-06-23 | 四川大学华西第二医院 | Gold-labeled kit with NCAM 1 as Testing index and its preparation method and application |
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|---|---|---|---|---|
| CN106885902A (en) * | 2017-03-03 | 2017-06-23 | 四川大学华西第二医院 | Gold-labeled kit with NCAM 1 as Testing index and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| ABDEL-NASSER KAWDE ET AL: "Moving Enzyme-Linked ImmunoSorbent Assay to the Point-of-Care Dry-Reagent Strip Biosensors", 《AMERICAN JOURNAL OF BIOMEDICAL SCIENCES》 * |
| SHAOJUN GUO ET AL.: "A Novel Urchinlike Gold/Platinum Hybrid Nanocatalyst with Controlled Size"，", 《THE JOURNAL OF PHYSICAL CHEMISTRY C》 * |
| ZHUANGQIANG GAO ET AL.: "Urchin-like(gold core)@(platinum shell)nanohybrids:A highly efficient peroxidase-mimetic systemfor in situ amplified colorimetric immunoassay", 《BIOSENSORS AND BIOELECTRONICS》 * |
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Application publication date: 20190618 |