SBIR-STTR Award

The long-term objective of this proposal is to further develop a photochemical signal amplification method (PSAM) for increasing the sensitivity of conventional enzyme-linked immunosorbent assays (ELISA) for determination of biomarkers for various neurodegenerative diseases, such as Alzheimer's Disease (AD).Immunoenzymatic methods such as ELISA are widely used in biology and medicine for drug screening, for detecting viruses (including coronaviruses), bacteria, and biomarkers, particularly cytokines and interleukins for cancer and immunological disorders, and biomarkers for neurodegenerative diseases. They are routinely used in manual, semi-automated, and automated systems where high volumes of tests are performed. However, in many cases the sensitivity of ELISA is inadequate. Some of the key biomarkers for early diagnosis of AD include three cerebrospinal fluid (CSF) biomarkers:amyloid ß-42 (Aß-42), which causes formation of oligomeric plaques, total tau (t-tau), and phosphorylated tau(p-tau) (which increases intracellular neurofibrillary tangles (NFTs). To date, there is no sensitive and cost-effective method for the quantification of these three proteins, hindering the diagnosis and progression monitoring of AD. Existing highly sensitive methods are cumbersome and expensive. Therefore, there is an increased necessity for a common ELISA platform to detect low levels of Aß-42, t-tau, p-tau and other biomarkers for various neurodegenerative diseases that is both inexpensive and simple enough to not require specialized laboratory equipment. In our Phase I project, we propose to further develop a very sensitive and inexpensive immunoassay platform for detection of biomarkers for various neurodegenerative diseases. The proposedtechnology, ELISA + PSAM, consists of two steps: (1) a conventional horseradish peroxidase (HRP)-mediatedassay (ELISA) in which a common chromogenic HRP substrate, is used and (2) a substrate solution containing the product of the enzymatic reaction is irradiated by visible light. Illumination of the sample leads to a DAP-catalyzed drastic increase in DAP concentration (autocatalytic photochemical reaction). The Specific aims of Phase I are: 1. To explore all variables affecting the performance of ELISA + PSAM assays for detection of Aß-42, t-tau and p-tau and optimize the performance of the assays. To develop a universal approach for transitioning from ELISA to ELISA + PSAM assays. 2. To validate the universal approach for optimization of ELISA + PSAM assays by using up to 3commercially available ELISA kits from different manufacturers for each analyte. This will include determination of sensitivity and reproducibility/precision of the assays. Narrative The development of the proposed method will allow using highly sensitive tests for detection of low concentrations of biomarkers for various neurodegenerative diseases. The performance of the developed tests will be improved significantly compared to conventional assays. Successful completion of these studies will be beneficial for public health and make available all the technology needed for a substantial business opportunity to license the technology and manufacture commercial products. Achievement ; Achievement Attainment ; Affect ; Alzheimer's Disease ; AD dementia ; Alzheimer ; Alzheimer Type Dementia ; Alzheimer disease ; Alzheimer sclerosis ; Alzheimer syndrome ; Alzheimer's ; Alzheimer's disease dementia ; Alzheimers Dementia ; Alzheimers disease ; Primary Senile Degenerative Dementia ; dementia of the Alzheimer type ; primary degenerative dementia ; senile dementia of the Alzheimer type ; Antibodies ; Award ; Bacteria ; Belief ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Biology ; Blood ; Blood Reticuloendothelial System ; Malignant Neoplasms ; Cancers ; Malignant Tumor ; malignancy ; neoplasm/cancer ; Cerebrospinal Fluid ; cerebral spinal fluid ; spinal fluid ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; ELISA ; Enzymes ; Enzyme Gene ; Horseradish Peroxidase ; Immunoassay ; Immune System Diseases ; Immune Diseases ; Immune Disorders ; Immune Dysfunction ; Immune System Disorder ; Immune System Dysfunction ; Immune System and Related Disorders ; Immunodeficiency and Immunosuppression Disorders ; Immunologic Diseases ; Immunological Diseases ; Immunological Dysfunction ; Immunological System Dysfunction ; Interleukins ; Lighting ; Illumination ; Manuals ; Medicine ; Methods ; Legal patent ; Patents ; Phenylenediamines ; Plasma ; Blood Plasma ; Plasma Serum ; Reticuloendothelial System, Serum, Plasma ; Proteins ; Public Health ; Reagent ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Technology ; Testing ; Time ; Virus ; cytokine ; Measures ; HIV Core Protein p24 ; HIV Major Core Protein p24 ; HIV Protein p24 ; HIV gag Gene Product p24 ; HIV gag Protein p24 ; HIV p24 Antigen ; HIV-1 Core Protein p24 ; HTLV-III p24 ; Neurofibrillary Tangles ; neurofibrillary degeneration ; neurofibrillary lesion ; neurofibrillary pathology ; tangle ; tau Proteins ; MT-bound tau ; microtubule bound tau ; microtubule-bound tau ; tau ; tau factor ; τ Proteins ; Businesses ; Mediating ; analytical method ; base ; Blood specimen ; Blood Sample ; Label ; improved ; Solid ; Phase ; Coronavirus ; Coronaviridae ; corona virus ; Serum ; Blood Serum ; European ; Licensing ; Visible Radiation ; Visible Light ; Visible Light Radiation ; tau-1 ; p-tau ; p-τ ; phospho-tau ; phospho-τ ; phosphorylated tau ; Complex ; Reaction ; System ; Degenerative Neurologic Diseases ; Degenerative Neurologic Disorders ; Nervous System Degenerative Diseases ; Neural Degenerative Diseases ; Neural degenerative Disorders ; Neurodegenerative Diseases ; Neurologic Degenerative Conditions ; degenerative diseases of motor and sensory neurons ; degenerative neurological diseases ; neurodegenerative illness ; Neurodegenerative Disorders ; interest ; Performance ; success ; novel technologies ; new technology ; Sampling ; Property ; Manufacturer ; Manufacturer Name ; Enzymatic Reaction ; Biochemical Reaction ; Detection ; Reproducibility ; Validation ; Monitor ; Process ; Development ; developmental ; cost ; design ; designing ; cost effective ; innovation ; innovate ; innovative ; Biological Markers ; bio-markers ; biologic marker ; biomarker ; Secure ; laboratory equipment ; lab equipment ; laboratory technology ; early detection biomarkers ; early biomarkers ; early detection markers ; Drug Screening ; Alzheimer's disease biomarker ; Alzheimer's biomarker ; Alzheimer's disease biological marker ; Alzheimer's biological marker ; Amyloid beta-42 ; A β-42 ; A β42 ; A-beta 42 ; A-beta42 ; Abeta-42 ; Abeta42 ; Amyloid beta42 ; Amyloid β-42 ; Amyloid β42 ; Amyloidβ-42 ; Amyloidβ42 ; Aβ-42 ; Aβ42 ; Alzheimer's disease diagnosis ; Alzheimer's diagnosis ; antibody detection ; antibody based detection ; detect antibodies ; detection test ; detection tests ; detection assay ; detection limit ; detection method ; detection procedure ; detection technique ; detection platform ; detection system ;

The long-term objective of this proposal is to further develop a photochemical signal amplification method(PSAM) for increasing the sensitivity of conventional enzyme-linked immunosorbent assays (ELISA) fordetermination of biomarkers for various neurodegenerative diseases, such as Alzheimer's Disease (AD).Immunoenzymatic methods such as ELISA are widely used in biology and medicine for drug screening, fordetecting viruses (including coronaviruses), bacteria, and biomarkers, particularly cytokines and interleukins forcancer and immunological disorders, and biomarkers for neurodegenerative diseases. They are routinely usedin manual, semi-automated, and automated systems where high volumes of tests are performed. However, inmany cases the sensitivity of ELISA is inadequate. Some of the key biomarkers for early diagnosis of AD include three cerebrospinal fluid (CSF) biomarkers:amyloid ß-42 (Aß-42), which causes formation of oligomeric plaques, total tau (t-tau), and phosphorylated tau(p-tau) (which increases intracellular neurofibrillary tangles (NFTs). To date, there is no sensitive and cost-effective method for the quantification of these three proteins, hindering the diagnosis and progressionmonitoring of AD. Existing highly sensitive methods are cumbersome and expensive. Therefore, there is an increasednecessity for a common ELISA platform to detect low levels of Aß-42, t-tau, p-tau and other biomarkers forvarious neurodegenerative diseases that is both inexpensive and simple enough to not require specializedlaboratory equipment. In our Phase II project, we propose to further develop a very sensitive and inexpensiveimmunoassay platform for detection of biomarkers for various neurodegenerative diseases. In particular, we will1) design and manufacture a pre-production PSAM illumination device; 2) design and develop a PSAM detectionsystem kit, ELISA + PSAM kits for detection of Aß-42, t-tau, p-tau, BDNF and proBDNF; 3) conduct extensiveperformance evaluation studies towards Research Use Only applications of the PSAM products.

Public Health Relevance Statement:
Narrative The development of the proposed method will allow using highly sensitive tests for detection of low concentrations of biomarkers for various neurodegenerative diseases. The performance of the developed tests will be improved significantly compared to conventional assays. Successful completion of these studies will be beneficial for public health and make available all the technology needed for a substantial business opportunity to license the technology and manufacture commercial products.

Project Terms:
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