Preparation method of modified porous plate

A porous plate and modification technology, which is applied in the field of medical device modification and nanomaterials, can solve the problems of complicated experimental process, complicated operation and high cost, and achieve the effect of simple process, low cost and easy process

Active Publication Date: 2013-08-07
BIOMODI BIOTECH SUZHOU CO LTD
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  • Abstract
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Problems solved by technology

[0005] (1) Ultraviolet irradiation, plasma treatment and other methods are usually used to treat the surface of the microtiter plate to improve its binding ability to antigens; but this method requires special equipment, high cost, and complicated operation. In addition, these methods The modified surface can only increase the amount of antigen binding to a certain extent, but cannot promote the full exposure of antigenic epitopes
[0006] (2) Although the bead type, tube type and magnetic ball ELSIA method can solve this problem, the whole experimental process is complicated, the cost is high, and special equipment is required

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  • Preparation method of modified porous plate
  • Preparation method of modified porous plate

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Experimental program
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Embodiment 1

[0027] (1) 0.3g potassium bicarbonate (molecular weight 100), 0.03g glucose (molecular weight 198), 30mg chloroauric acid powder were dissolved in 6mL deionized water, and the pH value was adjusted to 9.0 with sodium hydroxide solution.

[0028] (2) Pipette 100 μL of the working solution into the microplate, and then place the microplate in an oven at 25° C. for 6 hours. The reaction solution in the wells was discarded, and then rinsed with deionized water for 3 times to prepare an ELISA plate modified with gold nanoparticles aggregates.

[0029] (3) Dilute human antithrombin with carbonate buffer (pH=9.6) to obtain protein solutions with concentrations of 96ng / mL, 19.2g / mL, 3.84ng / mL, and 0.768ng / mL . The above-mentioned gradient solution was coated on the ordinary microplate and the modified microplate at the dosage of 100 μL per well, and incubated in an oven at 37°C for 2 hours. Discard the coating solution in the plate, wash it with plate washing solution for 3 times, t...

Embodiment 2

[0032] (1) 0.3g potassium bicarbonate (molecular weight 100), 0.03g glucose (molecular weight 198), 30mg chloroauric acid powder were dissolved in 6mL deionized water, and the pH value was adjusted to 9.0 with sodium hydroxide solution.

[0033] (2) Pipette 250 μL of the working solution into the microplate, and then place the microplate in an oven at 50° C. for 1 hour to react. The reaction solution in the wells was discarded, and then rinsed with deionized water for 3 times to prepare an ELISA plate modified with gold nanoparticles aggregates.

[0034](3) The human plasma was serially diluted with carbonate buffer (pH=9.6). 100 μL per well was coated on ordinary microplates and modified microplates, and incubated in an oven at 37°C for 2 hours. Discard the coating solution in the plate, wash it with plate washing solution for 3 times, then pat dry the residual liquid in the porous plate on absorbent paper, add 300 μL per well of blocking solution containing 1.5% bovine seru...

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Abstract

The present invention belongs to the field of modification of a medical apparatus and the field of a nano material, and particularly relates to a preparation method of a modified porous plate, which comprises the following steps: (1), preparing aqueous solution containing potassium bicarbonate, glucose and chloroauric acid at 0-25 DEG C, adjusting the pH of the solution to be 9.0-10.0 with alkaline solution, and cooling the solution; and (2) filling the solution obtained in Step (1) into holes of the porous plate with 50-250muL for each hole, heating the porous plate for 1-6 hour(s) at 25-50 DEG C, eliminating the solution in the holes, and washing the porous plate for twice to three times with deionized water, therefore, the stable porous plate collectively modified by gold nanoparticlesis obtained. The gold nanoparticles are produced through chemical reduction, and are attached to the surface of the porous plate through physical settlement to form a stable and uniform gold nanoparticle aggregation layer. Due to the large specific surface area and the unique three-dimensional structure of the gold nanoparticle aggregation layer, the protein binding capacity of the modified surface can be improved. Therefore, the modified porous plate can realize ultra-high-sensitivity enzyme-linked immunosorbent assay tests.

Description

technical field [0001] The invention belongs to the field of modification of medical equipment and the field of nanomaterials, and specifically relates to a method for modifying a nano-gold particle layer on the surface of an enzyme-labeled plate. Using a porous plate (including an enzyme-labeled plate) modified with a nano-gold particle layer can significantly improve the traditional Detection limit of ELISA. Background technique [0002] In the field of various immunodiagnostics based on solid-phase carriers, enzyme-linked immunosorbent assay (ELISA), as one of the most important disease diagnostic techniques, has the advantages of high sensitivity, simple operation, low cost, and strong adaptability, so it is widely used in scientific research and medical fields. [0003] However, for the early stages of many malignant diseases, the content of related characteristic proteins in the blood is extremely small, which is often lower than the detection limit of commercially av...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/535G01N33/68
Inventor 陈红周峰袁琳
Owner BIOMODI BIOTECH SUZHOU CO LTD
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