We propose to develop an inexpensive high-throughput means of selecting and identifying antibodies to both unmodified and modified protein, peptide, and non-peptide antigens. The specific goals of this proposal are to complete a proof-of-concept using aqueous droplets in an oil emulsion for identification and isolation of scFv antibodies. Compartmentalization of recombinant-phage-producing E.coli cells is used to individually query the ability of each scFv to bind to antigen-coated beads. We propose to develop and test these methods on a phosphopeptide epitope found on the protein Her2. The successful completion of this SBIR and subsequent Phase II goals will have considerable direct impact on the development of new therapeutic targets, and immunotherapeutic, prognostic, and diagnostic biomarkers.
Public Health Relevance: Gene arrays enabled great breakthroughs in our understanding of the underlying biological processes that occur in cells and tissues. These increased understandings have already led to new approaches to diagnosis and treatment of major diseases, including cancer, diabetes and Alzheimer's. We are now entering a post-genomic era where the need for proteomics tools is becoming increasingly important. The technical challenges for proteomic tools are far greater than those encountered during the genome project. This proposal describes the high-throughput manufacture of antibodies for measuring protein abundance. The successful completion of this SBIR and our combined future goals will have significant direct impact on the development of new therapeutic targets as well as immunotherapeutic, prognostic, and diagnostic biomarkers.
Public Health Relevance Statement: PROJECT NARRATIVE Gene arrays enabled great breakthroughs in our understanding of the underlying biological processes that occur in cells and tissues. These increased understandings have already led to new approaches to diagnosis and treatment of major diseases, including cancer, diabetes and Alzheimer's. We are now entering a post-genomic era where the need for proteomics tools is becoming increasingly important. The technical challenges for proteomic tools are far greater than those encountered during the genome project. This proposal describes the high-throughput manufacture of antibodies for measuring protein abundance. The successful completion of this SBIR and our combined future goals will have significant direct impact on the development of new therapeutic targets as well as immunotherapeutic, prognostic, and diagnostic biomarkers.
NIH Spending Category: Biotechnology; Genetics
Project Terms: ATGN; Address; Affinity; Alzheimer; Alzheimer disease; Alzheimer sclerosis; Alzheimer syndrome; Alzheimer's; Alzheimer's Disease; Alzheimers Dementia; Alzheimers disease; Antibodies; Antigenic Determinants; Antigens; Assay; Bacteria; Bacteriophage M13; Bacteriophages; Binding; Binding (Molecular Function); Binding Determinants; Bioassay; Biologic Assays; Biological Assay; Biological Function; Biological Process; Body Tissues; Cancers; Cells; Coliphage M13; Coupled; Cytofluorometry, Flow; Dementia, Alzheimer Type; Dementia, Primary Senile Degenerative; Dementia, Senile; Development; Diabetes Mellitus; Diagnosis; Diagnostic; Disease; Disorder; E coli; Emulsions; Enterobacteria phage M13; Epitopes; Escherichia coli; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Fluorescein; Fluoresceins; Free Radicals; Funding; Future; Gamma Globulin, 7S; Gene Expression; Genes; Genetic Polymorphism; Genome; Genomics; Goals; HRP; Horseradish Peroxidase; Human; Human, General; Hydrogen Oxide; IgG; Image; Immunoglobulin G; Immunotherapeutic agent; Incubated; Individual; Investigators; L-tyrosine, dihydrogen phosphate (ester); Label; Libraries; Life; Ligands; M13 Phage; Malignant Neoplasms; Malignant Tumor; Man (Taxonomy); Man, Modern; Measures; Methods; Microbeads; Microfluorometry, Flow; Microspheres; Molecular; Molecular Interaction; NCI; NCI Organization; National Cancer Institute; Oils; Peptides; Phage Display; Phages; Phase; Phosphopeptides; Phosphotyrosine; Polymorphism (Genetics); Polymorphism, Genetic; Post-Translational Modifications; Post-Translational Protein Processing; Posttranslational Modifications; Primary Senile Degenerative Dementia; Production; Protein Modification; Protein Modification, Post-Translational; Protein Processing, Post-Translational; Protein Processing, Posttranslational; Protein/Amino Acid Biochemistry, Post-Translational Modification; Proteins; Proteome; Proteomics; RNA Splicing; Reagent; Recombinants; Research Personnel; Researchers; SBIR; SBIRS (R43/44); STTR; Small Business Innovation Research; Small Business Innovation Research Grant; Small Business Technology Transfer Research; Sorting - Cell Movement; Splicing; System; System, LOINC Axis 4; Testing; Tissues; Two Hybrid; Tyrosine-O-phosphate; Variant; Variation; Water; Yeast One Hybrid System; Yeast One/Two-Hybrid System; Yeasts; aqueous; bacterial virus; biomarker; dementia of the Alzheimer type; diabetes; disease/disorder; flow cytophotometry; gene product; imaging; immunogen; immunologic preparation; immunotherapeutics; malignancy; neoplasm/cancer; new approaches; new therapeutic target; novel approaches; novel strategies; novel strategy; polymorphism; primary degenerative dementia; prognostic; public health relevance; senile dementia of the Alzheimer type; sorting; tool; yeast two hybrid system
