Biochip sensor surface carrying polyethylene glycolated nanoparticles

A biosensor and nanoparticle technology, applied in the direction of nanotechnology, nanotechnology, chemical instruments and methods, etc., can solve the problems of different sensitivity enhancement effects, inability to prevent non-specific adsorption of impurities, and low sensitivity, so as to suppress non-specificity Adsorption effect

Inactive Publication Date: 2005-07-27
JAPAN SCI & TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the system using the above-mentioned gold nanoparticles for improving SPR sensitivity, it was found that the sensitivity enhancement effect was different due to the distance between the gold nanoparticles and the surface of the gold thin film of the biosensor chip, or the binding form of the particles and the surface ( See eg N. Nath et al.)
Therefore, even if the above-mentioned system using gold nanoparticles and a biosensor chip is applied to the system described in U.S. Patent No. 5,763,191, the sensitivity may be low or non-specific adsorption of impurities may not be prevented.

Method used

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  • Biochip sensor surface carrying polyethylene glycolated nanoparticles
  • Biochip sensor surface carrying polyethylene glycolated nanoparticles
  • Biochip sensor surface carrying polyethylene glycolated nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0094] Preparation Example 1: Preparation method (one) of PEG-modified gold particles:

[0095] Polymer used: Acetal-PEG-SH (Mn=5000)

[0096]

[0097] Acetal-PEG-SH:HAuCl 4 =1 / 6:1 (molar ratio), add relative HAuCl to the aqueous solution mixed in the above ratio 4 10 times the molar amount of NaBH 4 , Colloidal gold adjusted by reduction method. Treat the terminal acetal group with hydrochloric acid at pH 2, reduce it to an aldehyde group, and then react with p-aminophenyl-β-D-lactopyranoside to obtain a colloidal gold aqueous solution modified by lactose-PEG-SH (average particle size : 8.7nm).

[0098] Acetal-PEG-SH was prepared as follows.

[0099]Add 20ml of distilled tetrahydrofuran (THF) and 0.2mmol (0.032ml) 3,3-diethoxyl-1-propanol initiator in the container replaced by argon, then add the equivalent of potassium naphthyl, and stir for 15 minutes , for metallization. Thereafter, 22.7 mmol (1, 135 ml) of ethylene oxide was added, stirred at room temperature fo...

preparation example 2

[0100] make 2.0 x 10 -2 One mmol (100 mg) of PEG-SS-Py was dissolved in 4 ml of distilled water, and 0.1 mmol (15.42 mg) of dithiothreitol was added in a 5-fold molar amount, and stirred at room temperature for 30 minutes. After the reaction, the polymer (hereinafter referred to as PEG5000) was recovered by washing with saturated brine, extracting with chloroform, and reprecipitating with ether. recycled polymer 1 The structure was confirmed by H-NMR, and the terminal SH group was quantified by reacting with 2-pyridyl disulfide (2-PDS). Preparation Example 2: Preparation method of PEG-modified gold particles (two)

[0101] Polymer used: Acetal-PEG-SH (Mn=3200)

[0102]

[0103] (1) Preparation of the polymer used

[0104] According to the reaction scheme I, using the initiator 3,3-diethoxy-1-propanol and the terminator methanesulfonyl chloride, a heterobifunctional PEG with acetal group and methanesulfonyl group was synthesized by anionic polymerization. Then react wit...

preparation example 3

[0116] Preparation Example 3: Preparation method of PEG-modified gold particles (Part 3)

[0117] Polymer used in this example (Acetal-PEG-PAMA):

[0118]

[0119] (According to above-mentioned people such as kataoka, the method described in Macromolecules, 1999, 32, 6892-6894 pages obtains. Use PEG Mw=5000g / mol, PAMA (poly[methacrylic acid (2-N, N-dimethylamino ) ethyl ester]) n=130, m=100) to produce PEGylated CdS semiconductor particles. 1 mL of 2.5 mg / mL auric acid chloride (HAuCl 4 ) aqueous solution and 5 mL of 6 mg / mL acetal-PEG / PAMA block copolymer aqueous solution (NH:Au=8:1) were mixed, and stirred at room temperature for 24 hours. The UV-vis spectrum was measured at predetermined time intervals, and it was confirmed that the peak at 540 nm originating from the gold particles gradually increased, and a colloidal gold particle (fine particle) dispersion was produced without adding a reducing agent. The solution was measured by light scattering (DLS: Dynamic Ligh...

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Abstract

The invention provides high sensitivity bioassay sensor systems in which non-specific adsorption of impurities such as, for example, proteins, in biological samples is inhibited. Polyethylene glycolated particles enclosing metal or semi-conductor which is in common with the sensor material are used for amplification.

Description

technical field [0001] The present invention relates to the technical field of bioassay, more specifically, relates to reducing or preventing non-specific adsorption or binding caused by impurities other than analytes contained in biological fluids, etc., and improving the detection of analytes Sensitivity biosensor system, and assay method using the biosensor system. Background technique [0002] As a method for detecting an analyte present in a biological sample, biosensors having various detection methods have been proposed. Among such biosensors, sensors using surface plasmon resonance (hereinafter also referred to as SPR) are sensitive to changes in the refractive index of the metal thin film and its vicinity (for example, refer to A. Szabo et al., Curr. Opin. Strnct. Biol. 5 (1995) pp. 699-705). SPR is suitable for in-situ observation of the process between the surface and the complex biological solution, and for example, without the use of labels, the data of the an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J38/10G01N33/543G01N33/551G01N33/553
CPCG01N33/54346G01N33/54393B82Y15/00G01N33/553G01N33/551B82Y30/00G01N33/543G01N33/545
Inventor 片冈一则长崎幸夫大塚英典内田胜美石井武彦铃木祐子秋山好嗣高江誓词
Owner JAPAN SCI & TECH CORP
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