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Method for quantitatively detecting oleylamine grafted polysuccinimide macromolecule nanometer drug carrier based on indirect competitive enzyme-linked immunosorbent assay

A technology of polysuccinimide and enzyme-linked immunosorbent immunoassay, which is applied in the direction of biological testing and material inspection products, can solve the problems of few researches on the quantitative detection of nanocarriers, and achieve the effects of low detection limit, high sensitivity and long shelf life

Active Publication Date: 2017-05-24
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It can be seen that there are few studies on the quantitative detection of nanocarriers in vivo

Method used

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  • Method for quantitatively detecting oleylamine grafted polysuccinimide macromolecule nanometer drug carrier based on indirect competitive enzyme-linked immunosorbent assay

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preparation example Construction

[0039] The preparation method of each solution that the present invention relates to is:

[0040] PBS buffer: weigh 8.0g NaCl, 0.1g KCl, NaH 2 PO 4 2H 2 O 0.29g, Na 2 HPO 4 12H 2 Dissolve O2.96g in distilled water and set the volume to 1000mL to obtain 0.01mol / L PBS buffer solution with pH=7.4;

[0041] PBST solution: add 500μL Tween-20 to 1000mL PBS, mix well;

[0042] Coating buffer CB: weigh Na 2 CO 3 1.59g, NaHCO 3 Dissolve 2.94g in distilled water and make up to 1000mL; you can get 0.05mol / L coating buffer solution with pH=9.6;

[0043] O-phenylenediamine substrate solution: weigh 1.85g Na 2 HPO 4 12H 2 O, 0.51g C 6 h 8 o 7 Dissolve in distilled water and set the volume to 50mL, weigh 4mg of o-phenylenediamine and dissolve in 10mL of the above solution, add 15μL of 30% H 2 o 2 ;

[0044] 1 wt% casein: Weigh 1 mg of casein and dissolve it in 1 mL of 0.01 mol / L PBS buffer solution with pH=7.4, and mix well.

Embodiment 1

[0046] A Quantitative Detection of PSI Based on Indirect Competitive ELISA OAm method, including the following steps:

[0047] a. Preparation of PSI OAm Coating antigens and immunogens specifically includes the following steps:

[0048] a-1. Hydrolyzed PSI OAm , to get the PSI after hydrolysis OAm solution;

[0049] With reference to the method in Nano Research, 2015, 8(6):1932-1943, a chloroform solution of oleylamine-grafted polysuccinimide was prepared, and the steps were as follows: take 32mL N, N-dimethylformaldehyde Heat the amide to 90°C, add 1.6g polysuccinimide and 2.17mL oleylamine, keep heating at 100°C for 5 hours, finally add 80mL methanol to precipitate, centrifuge, weigh the precipitate and disperse in 2-10mL chloroform , the solution concentration was adjusted to 60 mg / mL to obtain a chloroform solution of oleylamine grafted polysuccinimide.

[0050] Take 0.3mL of this solution and disperse it in 3mL 0.5mmol / L NaOH, ultrasonicate at 150W for 7min, stir th...

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Abstract

The invention discloses a method for quantitatively detectingan oleylamine grafted polysuccinimide(PSIOAm) macromolecule nanometer drug carrier based on an indirect competitive enzyme-linked immunosorbent assay. A secondary antibody is utilized, so that a detection signal is amplified, and the sensitivity of experimental analysis is improved; the aim of quantitatively detecting the PSIOAm is achieved through measuring an optical signal of a compound of a PSIOAm envelope antigen, a PSIOAm antibody which is the primary antibody and an HRP-marked goat anti-rabbit antibody which is the secondary antibody. The method is simple to operate, high in feasibility, high in sensitivity, low in detection limit, and capable of realizing high throughput detection.

Description

technical field [0001] The present invention relates to the quantitative detection of nanomaterials, in particular to a quantitative detection of oleylamine grafted polysuccinimide (PSI) based on an indirect competitive enzyme-linked immunosorbent assay OAm ) The method of polymer nano drug carrier. Background technique [0002] In the 1960s, chemists proposed that polymer materials could be applied in the field of biomedicine to synthesize a polymer drug carrier to improve the drug structure and improve drug efficacy. The polymer drug carrier itself does not have pharmacological effects, nor does it chemically react with the drug, but combines with the low-molecular drug in an appropriate way (hydrogen bond, ionic bond or complex form, mainly the former). High-molecular compounds are only used as delivery systems for low-molecular drugs, and it is still low-molecular drugs that really exert pharmacological effects. [0003] Polymer nano-drug carrier refers to a nano-scale...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/53
Inventor 张明翠曹以婷
Owner ANHUI NORMAL UNIV
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