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Liquid crystal based analyte detection

a liquid crystal and analyte technology, applied in the field of liquid crystal based analyte detection, can solve the problems of high cost, high complexity and/or complexity of methods with the highest sensitivity (real-time pcr, tissue culture, electron microscopy), and high cost, and achieve the effect of less sensitivity, high cost, and high sensitivity

Inactive Publication Date: 2005-03-24
PLATYPUS TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methods with the highest sensitivity (real-time PCR, tissue culture, electron microcopy) also involve the highest complexity and / or cost, require sophisticated equipment and facilities and require highly trained personnel.
Methods with less sensitivity (IFA, ELISA, dipstick methods), in practice, suffer from cross-reactivity problems, involve more hands-on time and / or are less adaptable to rapidly screening large numbers of samples.

Method used

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  • Liquid crystal based analyte detection
  • Liquid crystal based analyte detection
  • Liquid crystal based analyte detection

Examples

Experimental program
Comparison scheme
Effect test

example 1

Immobilization of Antibodies on Substrates

This example describes different methods for immobilizing antibodies on substrate. Five different immobilization strategies were evaluated:

1) HEXA: adsorption of Protein A, then the West Nile Virus monoclonal antibodies (WNV Mabs) onto a hydrophobic monolayer formed from CH3(CH2)15SH (HEXA) on the surface of a gold film. The surface was blocked with BSA after immobilization of the antibody.

2) SPDP: covalent attachment of WNV Mabs to a monolayer formed from 2-mercaptoethylamine (2-MEA) on a gold film by using the sulfhydryl-reactive (protein) and amine-reactive (monolayer) heterobifunctional cross-linker N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). The surface was blocked with BSA after immobilization of the antibody.

3) PMPI: covalent attachment of Ras polyclonal antibodies (Ras Pabs) to a monolayer formed from 11-mercaptoundecanol (11-MU) on a gold film by using a sulfhydryl-reactive (protein) and hydroxyl-reactive (monolayer...

example 2

Detection of West Nile Virus

The detection of viruses with liquid crystal assays in which the detection region comprises topographical features has been described in WO 01 / 61357. These types of surfaces in combination with liquid crystals were successfully used to report the presence of West Nile Virus (WNV) captured on the surface of such substrates. Surprisingly, however, it has now been found that the reporting mechanism does not require the topography on the surface. This unexpected outcome substantially simplifies the fabrication of substrates for detection of viruses using liquid crystals. As described below, it has been demonstrated that this reporting mechanism can be applied to different viruses.

Antibodies to WNV were deposited onto the surface of molded polyurethane replicas. The micromolded replicas had a pitch of 400 nm and a depth of 54 nm. A drop of aqueous solution containing WNV was deposited onto the surface of the polymeric replica. The solutions contained 108.4...

example 3

Optimization of Antibody Immobilization

Different methods of immobilizing the antibody were investigated to determine the procedure which would give the best results. Briefly, polyurethane substrates were functionalized with (a) 1 uM WNV monoclonal antibodies, (b) 5 uM WNV monoclonal antibodies, and (c) 1 mg / ml Protein A first, then 1 uM WNV monoclonal antibodies. All functionalized substrates were then incubated with the WNV stock. The results showed that substantially the same homeotropic response is observed when the polyurethane is functionalized with 1 uM or 5 uM antibody, and also when the substrate is first incubated with Protein A (molecule which correctly orients the antibody), and then functionalized with 1 uM antibody. These results indicate that a strong homeotropic response can be obtained with a lower concentration of antibody, with or without Protein A. The current method involves coating the entire substrate with 1 mg / ml Protein A, and subsequently immobilizing anti...

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Abstract

The present invention relates to the field of detection of viruses, and in particular to detection of viruses using a liquid crystal assay format. In the present invention, virus binding in a detection region is identified by changes in liquid crystal orientation caused by virus binding independent orientation caused by any topography associated with the detection region.

Description

FIELD OF THE INVENTION The present invention relates to the field of detection of analytes, and in particular to detection of viruses, cells, bacteria, lipid-membrane containing organisms, proteins, nucleic acids, carbohydrates and other biomolecules, organic molecules and inorganic molecules using a liquid crystal assay format. BACKGROUND OF THE INVENTION The detection of pathogen, protein, and nucleic acid targets in biological samples forms the basis of the multi-billion dollar in vitro diagnostic industry. Detection of protein and nucleic acid targets can be divided into diagnostic and research based markets. The diagnostic market includes the detection and identification of pathogens such as viruses and bacteria, the identification of various genetic markers, and the identification of markers associated with the presence of tumors. The research market includes the genomics and proteomics industries, which require analytical, drug discovery, and high-throughput screening techn...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/70G01NG01N33/543G01N33/569
CPCB82Y15/00G01N33/56983G01N33/54373B82Y30/00C12Q1/70Y02A50/30
Inventor ISRAEL, BARBARA A.ABBOTT, NICHOLASMURPHY, CHRISTOPHERHOLLISTER, KARLASOLTANINASSAB, SYRUSHANSMANN, DOUGACHARYA, BHARAT RAJ
Owner PLATYPUS TECH
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