A kind of immunogold probe modified by nucleic acid antibody and its preparation method and application
A nucleic acid antibody, immunogold technology, applied in instruments, measuring devices, scientific instruments, etc., can solve the problems of easy aggregation, poor specificity, unstable gold probe structure, etc., and achieves good dispersion, low cost, and good promotion. Foreground effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] 1. Materials and equipment
[0037] 1) 15nm gold nanoparticles were purchased from Dongna Biomaterials Co., Ltd.
[0038] 2) Alkyne-modified long-chain DNA, short-chain DNA, and sulfhydryl-modified antisense long-chain DNA were purchased from Takara Company, and their sequences were: —CHCH:TT TAT ATA GAG GAG TTTTT TTTTT, TTTTT TTTTT, —SH:CTC CTC TATATA AA TTTTT TTTTT TT
[0039] 3) Type 71 enterovirus antibody (ab36367) was purchased from Abcam;
[0040] 4) Copper sulfate (C112396), sodium erythorbate (S105024), THPTA (762342-100MG) (tris(3-hydroxypropyltriazolemethyl)amine), aminoguanidine hydrochloride (A111225) were purchased from Aladdin Biochemical Science and Technology Co., Ltd., the azide compound (R-M-3042) was purchased from Sinopharm Chemical Reagent Co., Ltd.;
[0041] 5) The dialysis bag was purchased from Spectrum;
[0042] 6) PBS (10X) was purchased from Jiangsu Biyuntian Biotechnology Co., Ltd.;
[0043] 7) The centrifuge was purchased from Eppendor...
Embodiment 2
[0059] The nucleic acid antibody-modified gold nanoprobe prepared in Example 1 was used for the experiment of capturing enterovirus particles of type 71 in the sample. When the prepared nucleic acid antibody-modified gold nanoprobe was mixed with the virus sample, The gold probe can specifically adsorb the virus, and the antibody modified on the surface of the probe binds to the virus, and then the detection sample is obtained by centrifugation and resuspension, such as image 3 The transmission electron microscope characterization diagram shown shows that almost every EV71 virus particle has a gold probe bound to the surface. It can be seen that the specificity of the gold nanoprobe modified by the nucleic acid antibody is very good, and the ratio of the number of the probe to the virus is 5: 1. The ratio of the number of probes to viruses can also range from 10:1-1:1, mainly because the number of probes is too large, which will cause false positive results, and the number of ...
Embodiment 3
[0061] The gold nanoprobe modified by the nucleic acid antibody prepared in Example 1 was subjected to an indirect enzyme-linked immunosorbent assay to verify its specificity. With fetal bovine serum albumin (Bsa) as a control group, 100 μl of EV71 was first added dropwise to a 96-well plate. Virus liquid samples and 2% Bsa dilution were combined in a 37°C incubator for 2 hours, then 200 μl of 4% skimmed milk powder was added dropwise to each well for blocking, and the wells were left to stand in a 37°C incubator for 1 hour, using TBST After the solution was washed 4 times, 50 μl of gold probe solution was added dropwise to each well, combined at 37°C for 1 hour, after the plate was washed 4 times with TBST, 100 μl of secondary antibody (purchased from Boaosen Biotechnology Co., Ltd.) was added to each well, and 37°C Combine for 1 hour, wash the plate 4 times with TBST, add TMB (3,3',5,5'-tetramethylbenzidine) substrate solution, leave at 37°C for 10 minutes, add 30 μl stop sol...
PUM
| Property | Measurement | Unit |
|---|---|---|
| particle diameter | aaaaa | aaaaa |
| particle diameter | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More