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Detection of biotargets using bioreceptor functionalized nanoparticles

a biomolecule and functional technology, applied in the field of biotarget detection systems and methods, can solve the problems of limited amplification effect of bar-code methods, inability to fully understand the effect of raman enhancement, and use of expensive biomolecules (dna) and rather sophisticated procedures and analytical instruments

Inactive Publication Date: 2012-03-08
UNIV OF CENT FLORIDA RES FOUND INC
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the bar-code method developed by Mirkin et al. has pushed the detection limit beyond the attomolar range, this method involves the use of expensive biomolecules (DNA) and rather sophisticated procedures and analytical instrumentation.
Moreover, the amplification effect of the bar-code method is limited to the number of DNA barcodes that can be attached to the nanoparticle surface.
Although surface enhanced Raman scattering has shown promising potential for label-free trace detection of biomolecules, the Raman enhancement effect is not well understood and further study is needed before a routine assay method can be developed from this effect.
For the DNA aptamer ligation method, the biggest problem is that PCR amplification is needed for the analysis and the type of proteins that can be detected using this method is relatively limited.

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  • Detection of biotargets using bioreceptor functionalized nanoparticles
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  • Detection of biotargets using bioreceptor functionalized nanoparticles

Examples

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example 1

[0059]Currently, GFAAS and ICP-MS are the two most sensitive techniques for ultra trace metal analysis. The detection limit of GFAAS was examined on gold nanoparticles. A citrate stabilized nanoparticles with an average core diameter around 35 nm were decomposed by a solution of I2 / KI, where the I2 oxidizes gold atoms from the nanoparticles into gold ions. This solution was then diluted into four concentrations ranging from 10−11 to 10−15 M. This nanoparticle concentration range corresponds to a gold ion concentration ranging from 10−5 to 10−9 M. The analysis of 20 μL samples demonstrated that the lowest concentration of gold ions that can be detected by GFAAS is at the order of 10−8 M as analyzed using a Perkin Elmer AAalyst 600 instrument, resulting from a gold nanoparticle concentration of 10−14 M (10 s of femtomolar). Therefore, the minimal number of gold nanoparticles that can be detected by GFAAS is approximately 10,000.

[0060]Similar analysis was conducted on silver nanopartic...

example 2

[0061]PSA is a 32 kDa single chain glycoprotein serine protease with a chymotrypsin like specificity produced by the secretory epithelium of the prostate gland. PSA that is normally secreted into the seminal fluid plays a functional role in the cleavage of the seminal vesicle proteins and the liquefaction of the seminal coagulum. Only low levels of PSA are normally present in the blood stream, and increasing serum concentrations indicate prostatic pathology, including benign prostatic hyperplasia and cancer of the prostate. Determination of PSA is now widely used for detection and management of patients with prostatic cancer and considered as the superior serological marker for cancer of the prostate. The atomic amplification assay approach was used to examine the detection of PSA and compared the results with standard ELISA assay.

[0062]A commercial ELISA kit for PSA assay along with a paired detector antibody was purchased from AnoGen, Inc. We conducted PSA assay using the ELISA ki...

example 3

[0066]A route to nanoparticle probes according to an embodiment of the invention is given in FIG. 4. Relatively monodispersed citrate-stabilized gold nanoparticles can be synthesized using the method of Turkevich et al. in aqueous solution with sizes ranging from 10-120 nm, by controlling the volume of the capping ligand, tri-sodium citrate. The synthesis of citrate-stabilized silver nanoparticles can be done using by using silver nitrate as in Lee et al. J. Phys. Chem. 86 (1982) 3391. After purification, citrate-stabilized gold or silver nanoparticles will be converted to thioctic acid-protected nanoparticles through the ligand place exchange reaction. Two thiol groups-terminated thioctic acid is used to provide structural stability to the nanoparticles. The carboxyl groups on the nanoparticles are then activated with EDC (N-(3-dimethylaminopropyl)-N′-ethyl-carbodiimide) and NHS (N-hydroxysuccinimide). Two types of oligo(ethylene glycol) amine ligands, one with two amino end groups...

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Abstract

A system for determining a presence of biomolecule targets includes a vessel holding a sample solution suspected of containing the biomolecule targets and a plurality of probes including bioreceptor functionalized metal nanoparticles including a plurality of metal atoms. The biomolecule targets when present in the sample solution bind to the plurality of probes. A separation device is for separating the plurality of probes into a first group of probes having the biomolecule targets bound thereto and a second group of probes not having the biomolecule targets bound thereto. A reagent is introduced via a port for breaking down the metal nanoparticles in the first group or second group of probes, wherein at least one signal moiety solution is formed. A device measures a parameter of the signal moiety solution.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a divisional application of U.S. Non-Provisional patent application Ser. No. 11 / 875,252 filed Oct. 19, 2007, entitled “Detection of Biotargets Using Bioreceptor Functionalized Nanoparticles”, which claims priority to U.S. Provisional Patent Application Ser. No. 60 / 862,125 filed Oct. 19, 2006, entitled “Ultra Sensitive Detection of Biotargets Using Ions, Discrete Radioactive Isotopes or Discrete Fluorescent Dye Molecules Derived from Bioreceptor Functionalized Nanoparticles” and U.S. Provisional Patent Application Ser. No. 60 / 887,889 filed Feb. 2, 2007, entitled “Ultra Sensitive Detection of Biotargets Using Ions, Discrete Radioactive Isotopes or Discrete Fluorescent Dye Molecules Derived from Bioreceptor Functionalized Nanoparticles”.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]The U.S. Government may have certain rights to the invention based on National Science Foundation Career Award DMR 0552...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N21/59G01N30/02C12M1/40C12M1/34G01N33/53G01N27/72B82Y15/00
CPCG01N33/54346
Inventor HUO, QUNLIU, XIONGDAI, QIU
Owner UNIV OF CENT FLORIDA RES FOUND INC
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