CN117890579A - Application of special photosensitive microsphere for homogeneous chemiluminescent system - Google Patents
Application of special photosensitive microsphere for homogeneous chemiluminescent system Download PDFInfo
- Publication number
- CN117890579A CN117890579A CN202311839550.8A CN202311839550A CN117890579A CN 117890579 A CN117890579 A CN 117890579A CN 202311839550 A CN202311839550 A CN 202311839550A CN 117890579 A CN117890579 A CN 117890579A
- Authority
- CN
- China
- Prior art keywords
- microsphere
- photosensitive
- special
- photosensitive microsphere
- microspheres
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000004005 microsphere Substances 0.000 title claims abstract description 101
- 238000000034 method Methods 0.000 claims abstract description 12
- 239000000427 antigen Substances 0.000 claims abstract description 6
- 102000036639 antigens Human genes 0.000 claims abstract description 6
- 108091007433 antigens Proteins 0.000 claims abstract description 6
- 230000008961 swelling Effects 0.000 claims abstract description 5
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 238000005516 engineering process Methods 0.000 claims description 8
- -1 phthalocyanine bis (trihexylsiloxy) silane compound Chemical class 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 108010090804 Streptavidin Proteins 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 238000001179 sorption measurement Methods 0.000 claims description 4
- 238000004020 luminiscence type Methods 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- IEQIEDJGQAUEQZ-UHFFFAOYSA-N phthalocyanine Chemical compound N1C(N=C2C3=CC=CC=C3C(N=C3C4=CC=CC=C4C(=N4)N3)=N2)=C(C=CC=C2)C2=C1N=C1C2=CC=CC=C2C4=N1 IEQIEDJGQAUEQZ-UHFFFAOYSA-N 0.000 claims description 2
- 230000009870 specific binding Effects 0.000 claims 2
- 239000011248 coating agent Substances 0.000 claims 1
- 238000000576 coating method Methods 0.000 claims 1
- 239000002131 composite material Substances 0.000 claims 1
- 230000008878 coupling Effects 0.000 claims 1
- 238000010168 coupling process Methods 0.000 claims 1
- 238000005859 coupling reaction Methods 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 claims 1
- 239000002086 nanomaterial Substances 0.000 claims 1
- 239000002861 polymer material Substances 0.000 claims 1
- 239000002994 raw material Substances 0.000 claims 1
- 108010074051 C-Reactive Protein Proteins 0.000 abstract description 21
- 102100032752 C-reactive protein Human genes 0.000 abstract description 21
- 238000001514 detection method Methods 0.000 abstract description 20
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 19
- 238000012360 testing method Methods 0.000 abstract description 6
- 230000005284 excitation Effects 0.000 abstract description 2
- 125000000524 functional group Chemical group 0.000 abstract description 2
- 230000001900 immune effect Effects 0.000 abstract 1
- 238000012986 modification Methods 0.000 abstract 1
- 230000004048 modification Effects 0.000 abstract 1
- 238000005457 optimization Methods 0.000 abstract 1
- 238000002156 mixing Methods 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 239000007987 MES buffer Substances 0.000 description 11
- 238000005406 washing Methods 0.000 description 11
- 239000006228 supernatant Substances 0.000 description 10
- 239000002105 nanoparticle Substances 0.000 description 9
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 6
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 4
- 230000008105 immune reaction Effects 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 239000006194 liquid suspension Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 229910052693 Europium Inorganic materials 0.000 description 2
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- OGPBJKLSAFTDLK-UHFFFAOYSA-N europium atom Chemical compound [Eu] OGPBJKLSAFTDLK-UHFFFAOYSA-N 0.000 description 2
- 238000003018 immunoassay Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- BZYUMXXOAYSFOW-UHFFFAOYSA-N 2,3-dimethylthiophene Chemical compound CC=1C=CSC=1C BZYUMXXOAYSFOW-UHFFFAOYSA-N 0.000 description 1
- 238000012815 AlphaLISA Methods 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 238000003016 alphascreen Methods 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000005465 channeling Effects 0.000 description 1
- 238000002038 chemiluminescence detection Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000008073 immune recognition Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000000504 luminescence detection Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- JACPFCQFVIAGDN-UHFFFAOYSA-M sipc iv Chemical compound [OH-].[Si+4].CN(C)CCC[Si](C)(C)[O-].C=1C=CC=C(C(N=C2[N-]C(C3=CC=CC=C32)=N2)=N3)C=1C3=CC([C]1C=CC=CC1=1)=NC=1N=C1[C]3C=CC=CC3=C2[N-]1 JACPFCQFVIAGDN-UHFFFAOYSA-M 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention relates to an application of a special photosensitive microsphere for a homogeneous chemiluminescent system, which comprises the following steps: first, the photosensitive microsphere needs to have dye which can accept 680nm light excitation; at the same time, the microsphere surface must be modifiable, allowing modification of specific functional groups to couple with the antibody, and then recognize the antigen to be tested, forming an "immunological binding" complex. The special photosensitive microsphere for the homogeneous chemiluminescence system is prepared by adopting a high-temperature swelling method through a series of condition optimization, and the performance of the photosensitive microsphere is verified by preparing a C-reactive protein (CRP) detection reagent for testing.
Description
Technical Field
The invention belongs to the technical field of chemiluminescence detection, and particularly relates to application of a special photosensitive microsphere for a homogeneous chemiluminescence system. This technique relies on specific luminescent microspheres that are capable of generating a light signal under specific conditions, thereby enabling minute concentrations of biomolecules to be detected and quantified.
Background
The current common homogeneous chemiluminescent immunoassay technology in the market is a novel chemiluminescent assay technology developed based on LOCI (Luminescent Oxygen Channeling Assay) technology, and representative products are the AlphaScreen and AlphaLISA detection reagents of Perkinelmer company. Two types of microspheres are included in such reagents: one is a photosensitive microsphere containing a compound such as a phthalocyanine molecule that can generate singlet oxygen under light of a specific wavelength; one is a luminescent microsphere containing a fluorescent material such as dimethylthiophene and europium that can react with singlet oxygen. The photosensitive microsphere can generate a large amount of singlet oxygen molecules under the irradiation of laser with the wavelength of about 680 nm. The half-life of singlet oxygen is only 4us, the decay is fast in an aqueous medium, and the singlet oxygen can be transmitted for a distance of about 200nm at maximum. If the luminescent microsphere can approach the photosensitive microsphere in a specific recognition mode such as immune recognition, fluorescent substances in the luminescent microsphere can rapidly react with singlet oxygen generated near the photosensitive microsphere, and europium element is excited by energy transfer to emit fluorescent signals with the wavelength of about 610 nm. Therefore, when the photosensitive microsphere and the luminous microsphere are coated with the antibody, the antigen or other specifically recognized molecules, the distance between the photosensitive microsphere and the luminous microsphere can be less than 200nm through bridging with the corresponding to-be-detected object in the detection sample. Irradiation of such a mixture with a laser light having a wavelength of 680nm will produce a fluorescence signal at 610nm, the signal intensity of which is linearly dependent on the amount of analyte in the test sample over a range. The concentration of the object to be detected in the detection sample in the sample can be quantitatively detected by detecting the intensity of the fluorescent signal and then comparing the fluorescent signal with the calibration curve.
Compared with the traditional homogeneous immunoassay technology, the alpha LISA technology has the following advantages: the core of Alpha-LISA technology is photosensitive microsphere and luminescent microsphere, and the surfaces of the two microspheres must have modifiable properties according to different requirements. The nanometer microsphere is adopted, the surface performance is good, different functional groups can be modified, the microsphere shape is uniform and stable, the dispersibility is good, and aggregation is not easy. The immune reaction is a homogeneous reaction mode, the photosensitive microspheres and the luminous microspheres which wrap the antibody or antigen are uniformly distributed in a reaction system, and the two microspheres move freely under the action of an object to be detected to complete the immune reaction, so that one-step detection is realized, and the immune reaction time of light-activated chemiluminescence is shortened. The whole reaction process does not need to clean and separate the uncombined sample and reagent, thereby realizing the cleaning-free detection, reducing the systematic error of the reaction and improving the accuracy of the immune reaction. With the application of the nanoscale microsphere and the use of a biotin-streptavidin signal amplification system, the signal transmission process of four reactions of excitation light, high-energy oxygen ions, a light-emitting ball and optical signals can amplify and rapidly emit light, and the sensitivity of measurement is ensured. The whole process of energy (light) generation, transmission and amplification is very stable and is not easily affected by other factors.
Disclosure of Invention
Conventional photosensitive microspheres have some limitations such as relatively poor sensitivity and linearity, which limit their use in more complex or low concentration sample detection. In order to solve the limitations, the defects of the traditional photosensitive microspheres in sensitivity and linearity are overcome, and the application of the traditional photosensitive microspheres in detection of more complex or low-concentration samples is realized. The experiment successfully prepares photosensitive microsphere nanoparticles by a high-temperature swelling adsorption method, and the streptavidin is successfully connected by carboxyl amino reaction on the basis. The nano particles can be kept in a suspension state in an aqueous solution, namely, a stable colloid emulsion state is kept in a detection system, so that uniform detection is realized. The successful preparation of the photosensitive microsphere nanoparticles lays a foundation for the establishment of a subsequent homogeneous phase luminescence detection system.
The invention provides a preparation method of a special photosensitive microsphere for a homogeneous chemiluminescent system, which comprises the following steps:
(1) Dissolving a phthalocyanine bis (trihexylsiloxy) silane compound in benzyl alcohol to prepare 10 mg/mL;
(2) Putting 175-nm carboxyl polystyrene microsphere aqueous solution with solid content of 50% and 200 mu L into a 5-mL brown penicillin bottle, putting into a magnetic stirring table, adding 1-mL glycol, magnetically stirring for 3 min to uniformly mix, then dripping 800 mu L of 10-mg/mL silicon phthalocyanine solution into the carboxyl modified polystyrene microsphere solution by using a liquid transfer gun, and magnetically stirring at 60 ℃ while dripping;
(3) After the dripping is finished, placing the brown penicillin bottle into an oil bath pot for reaction for 10 min, wherein the reaction condition is 110 ℃,1000 r/min;
(4) Stopping stirring after the oil bath reaction is completed, and cooling to room temperature; after cooling, adding absolute ethyl alcohol with equal volume and different concentration gradients, centrifuging for 30 min at 14000 rpm and 4 ℃, washing for several times until the supernatant after centrifugation is colorless, washing with ultrapure water for the last time, re-suspending the photosensitive nanoparticles in the ultrapure water, and preserving at 4 ℃ in a dark place;
(5) Thoroughly mixing the self-made photosensitive nanoparticles by a vortex mixer, and transferring the mixture into a brown centrifuge tube with the concentration of 0.5-mL;
(6) Placing the brown centrifuge tube into a centrifuge, centrifuging at 16000 rpm for 30 min at 4deg.C, and removing supernatant;
(7) Adding 1 mL connection buffer solution (0.05M MES,pH 6.0), and fully and uniformly mixing the photosensitive nanoparticles by using a vortex oscillator;
(8) Adding 25 mu L of 10 mg/mL NHS and 40 mu L of 10 mg/mL EDC, and fully and uniformly mixing the photosensitive nanoparticles by using a vortex oscillator;
(9) Using a sample rotary mixer, rotating at room temperature for 30 min;
(10) Repeating the step (6), adding 1 mL connection buffer solution, and fully and uniformly mixing the suspension microspheres by using a vortex mixer; discarding the supernatant, repeating the step (6), and discarding the supernatant;
(11) Adding 0.5 mg SA (dissolving streptavidin in 0.05M MES,pH 6.0 binding buffer), mixing thoroughly with vortex mixer, adding into the above photosensitive nanoparticle, and suspending by ultrasonic method; vortex with sample mixer overnight at room temperature;
(12) Add 200. Mu.L of blocking solution (0.05M MES, 2% BSA,100 mM ethanolamine), block 2 h, then repeat step (6);
(13) Washing (0.025M Tris-HCl, 0.0% Proclin (TM) -300, pH 7.8) microspheres with 1 mL washing solution, fully mixing the suspended microspheres with a vortex mixer, and repeating the step (6);
(14) Repeating the step (13), centrifugally washing four times to preserve the liquid suspension microsphere, and preserving at 2-8 ℃.
The feasibility of the special photosensitive microsphere is verified by measuring a CRP sample by using a homogeneous chemiluminescence method after the photosensitive microsphere is prepared. The main detection steps are as follows:
Tris-HCl is used as a detection buffer solution, and biotinylated-ab, luminescent microspheres and streptavidin photosensitive microspheres are used as detection reagents;
adding the biotinylated antibody, a sample to be tested (standard substance) and the luminescent microspheres into a microplate in a dark place, and reacting for 10 min at 37 ℃ in a dark place;
then adding the photosensitive microsphere in a dark place, reacting for 5 min at 37 ℃, reading the optical signal value and recording.
Drawings
FIG. 1 is a graph showing the comparison of the results of two types of microspheres, namely, the test results of preparing a C-reactive protein (CRP) detection reagent from each of a specially-made photosensitive microsphere and a commercially-available common photosensitive microsphere.
Examples
Example 1
The C-reactive protein (CRP) detection reagent was prepared separately using a custom made photosensitive microsphere and a commercially available common photosensitive microsphere, respectively.
The preparation method of the C-reactive protein (CRP) detection reagent by using the special photosensitive microsphere comprises the following steps:
(1) thoroughly mixing the self-made photosensitive microspheres by a vortex mixer, and transferring the mixture into a brown centrifuge tube with the concentration of 0.5 to mL; (2) placing the brown centrifuge tube into a centrifuge, centrifuging at 16000 rpm for 30 min at 4deg.C, and removing supernatant; (3) adding MES buffer solution with the concentration of 1 mL of 0.05M and the pH of 6.0, and fully and uniformly mixing the photosensitive nanoparticles by using a vortex oscillator; (4) add 25. Mu.L of 20 mg/mL NHS and 40. Mu.L of 20 mg/mL EDC and mix the photosensitive microspheres thoroughly with vortex shaker; (5) using a sample rotary mixer, rotating at room temperature for 30 min; (6) repeating the step (2), adding MES buffer solution with the concentration of 1 mL of 0.05M and the pH of 6.0, and fully and uniformly mixing the suspension microspheres by using a vortex mixer; discarding the supernatant, repeating the step (2), and discarding the supernatant; (7) adding 0.5 mg SA (dissolved with MES buffer solution with concentration of 0.05M and pH of 6.0), mixing thoroughly with vortex mixer, adding into the above photosensitive microsphere, and suspending by ultrasonic method; vortex with sample mixer overnight at room temperature; (8) add 200. Mu.L of blocking solution (0.05M MES, 2% BSA,100 mM ethanolamine), block 2 h, then repeat step (2); (9) washing the microspheres with 1 mL washing solution (0.025M Tris-HCl, 0.0% Proclin (TM) -300, pH 7.8), mixing the microspheres with a vortex mixer, and repeating the step (2); repeating the step (9), and centrifugally washing four times to preserve the liquid suspension microsphere at 2-8 ℃.
The preparation method of the C-reactive protein (CRP) detection reagent by using the commercially available common photosensitive microspheres is as follows:
(1) commercially available photosensitive microspheres were thoroughly mixed with a vortex mixer and transferred to a 0.5 mL to brown centrifuge tube; (2) placing the brown centrifuge tube into a centrifuge, centrifuging at 16000 rpm for 30 min at 4deg.C, and removing supernatant; (3) adding MES buffer solution with the concentration of 1 mL of 0.05M and the pH of 6.0, and fully and uniformly mixing the photosensitive microspheres by using a vortex oscillator; (4) add 25. Mu.L of 20 mg/mL NHS and 40. Mu.L of 20 mg/mL EDC and mix the photosensitive microspheres thoroughly with vortex shaker; (5) using a sample rotary mixer, rotating at room temperature for 30 min; (6) repeating the step (2), adding MES buffer solution with the concentration of 1 mL of 0.05M and the pH of 6.0, and fully and uniformly mixing the suspension microspheres by using a vortex mixer; discarding the supernatant, repeating the step (2), and discarding the supernatant; (7) adding 0.5 mg SA (dissolved with MES buffer solution with concentration of 0.05M and pH of 6.0), mixing thoroughly with vortex mixer, adding into the above photosensitive microsphere, and suspending by ultrasonic method; vortex with sample mixer overnight at room temperature; (8) add 200. Mu.L of blocking solution (0.05M MES, 2% BSA,100 mM ethanolamine), block 2 h, then repeat step (2); (9) washing the microspheres with 1 mL washing solution (0.025M Tris-HCl, 0.0% Proclin (TM) -300, pH 7.8), mixing the microspheres with a vortex mixer, and repeating the step (2); repeating the step (9), and centrifugally washing four times to preserve the liquid suspension microsphere at 2-8 ℃.
Example 2
The special photosensitive microsphere and the commercial common photosensitive microsphere are respectively used for preparing the test and comparison of the C-reactive protein (CRP) detection reagent.
The C-reactive protein (CRP) antigen is prepared into 0.5 mL split-charging for each bottle with serial concentration of 0 ug/mL, 3.91 ug/mL, 7.81 ug/mL, 15.63ug/mL, 31.25 ug/mL, 62.5 ug/mL, 125 ug/mL, 250 ug/mL and 500 ug/mL by using standard substance buffer solution, and is preserved at 4 ℃ for standby. Standard buffer: tris-HCl (20 mM,pH 7.5, 150 mM NaCl,2 mM CaCl2)
Samples of 9 concentrations of the above C-reactive protein (CRP) antigen diluted were simultaneously detected using the C-reactive protein (CRP) detection reagents of the two photosensitive microspheres prepared in example 1, and the luminescence value of each sample was recorded, and the detection data are shown in table 1. The test results are plotted against the graph (fig. 1) according to the data in table 1.
Table 1 test data
| Concentration of sample (ug/mL) | Luminous mean value RLU (Special photosensitive microsphere) | Luminous mean value RLU (common photosensitive microsphere) |
| 0 | 365 | 475 |
| 3.91 | 963 | 855 |
| 7.81 | 1830 | 1496 |
| 15.63 | 3476 | 2544 |
| 31.25 | 6605 | 4299 |
| 62.5 | 12616 | 8125 |
| 125 | 23970 | 15030 |
| 250 | 46023 | 27506 |
| 500 | 87444 | 46485 |
Analysis of results: in the concentration range of 0-500 ug/mL of C Reactive Protein (CRP), the correlation coefficient of the reagent prepared by specially-made photosensitive microspheres is R 2 0.9992, higher than the correlation coefficient R of the reagent prepared by common photosensitive microspheres 2 = 0.9915. Under the same sample concentration, the luminous value of the reagent prepared by the special photosensitive microsphere is higher than that of the reagent prepared by the common photosensitive microsphere, and the advantages of low-end sensitivity and high-end linearity are reflected. The comprehensive comparison can be carried out, and the performance of the reagent prepared by the special photosensitive microsphere is obviously better than that of the reagent prepared by the common photosensitive microsphere.
Conclusion: the special photosensitive microsphere can be used in homogeneous chemiluminescence technology, and the performance of the reagent prepared by the special photosensitive microsphere is obviously superior to that of the reagent prepared by the common photosensitive microsphere.
Claims (8)
1. An application of a special photosensitive microsphere for a homogeneous chemiluminescent system, which is characterized in that: the special photosensitive microsphere adopts a high-temperature swelling adsorption method, takes a phthalocyanine bis (trihexylsiloxy) silane compound and carboxylated polystyrene microsphere as raw materials, and then is prepared by coupling streptavidin through carboxyl amino reaction.
2. The tailored photosensitive microsphere according to claim 1, wherein the coating is performed using a bis (trihexylsiloxy) silane compound of phthalocyanine.
3. The tailored photosensitive microsphere according to claim 1-2, wherein the microsphere is a carboxylated polystyrene microsphere.
4. A tailored photosensitive microsphere according to claims 1-3, having a microsphere diameter of less than 200nm.
5. The special photosensitive microsphere according to claim 1-4, wherein the microsphere can receive 680 and nm light-excited dye, and the microsphere surface can be modified.
6. The specialized photosensitive microsphere of claims 1-5, wherein the microsphere enhances the luminescence properties of a homogeneous chemiluminescent system after being dispersed therein.
7. The special photosensitive microsphere according to claim 1-6, wherein the preparation method is a high-temperature swelling adsorption method, and the method is a material preparation technology which is generally used for manufacturing high polymer materials, nano materials or composite materials with specific functions or characteristics. The main steps include swelling, adsorption, curing and treatment.
8. The special photosensitive microsphere according to claim 1-7, wherein the microsphere surface is coated with antibodies, antigens or other specific binding molecules capable of specific binding to the sample to be tested.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311839550.8A CN117890579A (en) | 2023-12-29 | 2023-12-29 | Application of special photosensitive microsphere for homogeneous chemiluminescent system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311839550.8A CN117890579A (en) | 2023-12-29 | 2023-12-29 | Application of special photosensitive microsphere for homogeneous chemiluminescent system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117890579A true CN117890579A (en) | 2024-04-16 |
Family
ID=90645150
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311839550.8A Pending CN117890579A (en) | 2023-12-29 | 2023-12-29 | Application of special photosensitive microsphere for homogeneous chemiluminescent system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117890579A (en) |
-
2023
- 2023-12-29 CN CN202311839550.8A patent/CN117890579A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110982512B (en) | Co-doped fluorescent microsphere of multiple near infrared II-region fluorescent dyes, preparation and application thereof | |
| EP3839485B1 (en) | Microsphere composition for chemiluminescence analysis and use thereof | |
| CN116577319A (en) | Microsphere composition for chemiluminescence detection and application thereof | |
| CN110736739A (en) | homogeneous phase chemiluminescence detection kit and application thereof | |
| CN110736735A (en) | homogeneous phase chemiluminescence detection kit and application thereof | |
| JP2014163758A (en) | Cellulose particulate containing fluorochrome compound | |
| CN114752374B (en) | Preparation method of microsphere composition used for homogeneous phase chemiluminescence immunoassay technology | |
| CN110736736A (en) | homogeneous phase chemiluminescence POCT detection method and device using same | |
| CN112557667A (en) | C-reactive protein high-sensitivity immunoassay method based on double-color quantum dot ratio fluorescent probe | |
| CN117890579A (en) | Application of special photosensitive microsphere for homogeneous chemiluminescent system | |
| CN112730338A (en) | Ag @ Au-based porous structure dual-signal nano amplification probe and SPR immunodetection method thereof | |
| CN116559151A (en) | Microsphere composition for chemiluminescence detection and application thereof | |
| CN112114131A (en) | Homogeneous phase chemiluminescence detection method and application thereof | |
| CN113125704A (en) | Homogeneous phase chemiluminescence assay kit and application thereof | |
| CN112114130A (en) | Receptor reagent for homogeneous phase chemiluminescence detection and application thereof | |
| CN113125706A (en) | Competitive homogeneous phase chemiluminescence assay kit and application thereof | |
| CN116183909B (en) | Immunodetection method based on aggregation-induced emission microsphere and magnetic separation technology | |
| CN111122843A (en) | Homogeneous phase chemiluminescence detection kit | |
| CN113125703B (en) | Myoglobin homogeneous detection kit and application thereof | |
| CN117491648B (en) | Human body autoantibody detection material and preparation method thereof | |
| CN116027019B (en) | Additive for chemiluminescence immunoassay and application thereof | |
| CN113125705B (en) | Myoglobin homogeneous detection kit and application thereof | |
| CN116482352A (en) | MagLOCI composite microsphere and application thereof | |
| CN109632920B (en) | Preparation method of electrochemical signal marking material | |
| CN118311022A (en) | Light-activated chemiluminescence analysis method and application thereof in determination kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication |