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Assay reagents for a neurogranin diagnostic kit

a neurogranin and kit technology, applied in the field of biomarkers, can solve the problems of complex brain injuries, multiple severe clinical outcomes, and difficult detection of subclinical brain injuries, and children with sickle cell disease are at high risk for subclinical brain injuries

Inactive Publication Date: 2014-05-22
THE JOHN HOPKINS UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is based on the development of antibodies and antibody fragments that specifically bind to neurogranin, a protein involved in neurological disorders. The invention provides isolated antibodies that recognize different parts of neurogranin, including amino acids 1-78, amino acids 55-78, and a specific fragment of SEQ ID NO:4. These antibodies can be used to detect and measure neurogranin in various assays, such as ELISA, competitive-binding assay, radioimmunological assay, and fluorescence-activated cell sorting (FACS) assay. The invention also provides polynucleotide aptamers that specifically bind to neurogranin, allowing for the development of kits for detecting and measuring neurogranin. Overall, the invention provides tools for better understanding and diagnosing neurological disorders associated with neurogranin.

Problems solved by technology

Brain injuries are complex and can have multiple severe clinical outcomes.
Moreover, detection of subclinical brain injury is difficult, especially in children and neonates with birth-related injury.
In addition, children with sickle cell disease are at high risk for subclinical brain injury.
Untreated subclinical brain injuries in children can progress to overt stroke, neurological damage, learning problems and memory loss.
Unfortunately, clinical tools such as physical exam, and imaging (CT Scan or MRI) are subjective, not widely available, not sensitive or specific enough and or too costly to identify the infant, child or adult with brain injury.

Method used

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  • Assay reagents for a neurogranin diagnostic kit
  • Assay reagents for a neurogranin diagnostic kit
  • Assay reagents for a neurogranin diagnostic kit

Examples

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example 1

Development of Neurogranin Assay

[0100]The Identification of Human NRGN Specific Aptamers.

[0101]Systematic Evolution of Ligands by EXponential enrichment (SELEX) procedure was used to identify the human NRGN specific aptamers. Briefly, the specific aptamer was selected from a pool of single strand RNA by filter immobilization. The RNA-NRGN target complex can bind to nitrocellulose filter, and free RNA went through filtration. The specific aptamer was recovered from the filter and PCR amplified. After several round of selection, the specific aptamer with highest affinity with NRGN was enriched and sequencing identified.

[0102]Prepare the Library for Selection.

[0103]Single strand DNA oligo pool was chemically synthesized. The DNA oligo has 40mer random central core, which flanked by 2 constant sequences, and the library can be amplified by a pair of primers which target the 5′ and 3′ conserve ends of the library. The sequence of the library is 5′-TCTCGGATCCTCAGCGAG TCGTCTG (N40) CCGCATC...

example 2

Proteomic Study to Identify Brain Proteins Reveals Elevations of Neurogranin in Children with Sickle Cell Disease

[0146]Silent cerebral infarct (SCI) is the most common form of neurologic injury in sickle cell disease. It is associated with decreased neurocognitive function, and increased risk for progressive injury, including stroke and new or progressive lesions. SCI is defined as any ischemic lesion visible on multiple T2-weighted magnetic resonance images (MRI) that is not associated with a history or physical exam suggestive of a focal neurologic deficit. To date, a noninvasive, unbiased laboratory test for SCI does not exist, and the ability to identify children with SCD who are at risk for SCI remains limited. Furthermore, the diagnosis of SCI is made with surveillance MRI, which is costly and not done routinely at many institutions. As a result patients are often diagnosed after they have experienced SCI-related impairments.

[0147]The identification of plasma brain proteins th...

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Abstract

The present invention relates to the field of biomarkers. More specifically, the present invention relates to assay reagents useful in detecting neurogranin. In a specific embodiment, the present invention provides an isolated antibody or fragment thereof that specifically binds to neurogranin. In another embodiment, the present invention provides a polynucleotide aptamer that specifically binds neurogranin.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 485,375, filed May 12, 2011, which is incorporated herein by reference in its entirety.STATEMENT OF GOVERNMENTAL INTEREST[0002]This invention was made with U.S. government support under grant no. NHLBI 1 R01 HL091759-02, and NHLBI 5U54HL090515-02. The U.S. government has certain rights in the invention.FIELD OF THE INVENTION[0003]The present invention relates to the field of biomarkers. More specifically, the present invention relates to assay reagents useful in detecting biomarkers.INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY[0004]This application contains a sequence listing. It has been submitted electronically via EFS-Web as an ASCII text file entitled “P11546-02_Sequence_Listing_ST25.txt.” The sequence listing is 3,146 bytes in size, and was created on May 10, 2012. It is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVE...

Claims

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Application Information

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IPC IPC(8): G01N33/68C12Q1/68
CPCC07K16/18G01N33/566C12N15/115C12N2310/16C12Q1/68G01N33/6896C07K2317/30G01N2333/47G01N2800/28
Inventor EVERETT, ALLEN DALEYANG, JUNFU, ZONGMING
Owner THE JOHN HOPKINS UNIV SCHOOL OF MEDICINE