SBIR-STTR Award

We propose to develop a rapid, low cost and high-throughput method for generating recombinant engineered antibodies with improved properties for proteomic and immunoassay applications. Engineered antibody constructs such as recombinant Fab fragments and chimeric antibodies will be generated and expressed by directly modifying the antibody genes in antibody-producing hybridoma cell lines, thereby eliminating the need for gene cloning and transfection. In this Phase I SBIR, we will demonstrate feasibility by modifying ten (10) hybridoma cell lines that produce antibodies used in MSD's commercial immunoassay kits. We will demonstrate that we can modify the cell lines so that they express their antibodies as recombinant Fab fragments or recombinant chimeras comprising Fab fragments linked to protein domains that enable site- specific labeling. The use of antibodies from established kits will make it straightforward to directly compare the assay performance of the new antibody constructs to the original full length antibodies. The long term goal of the program is to take advantage of beneficial properties of the chimeric constructs - lower non-specific interactions and the ability to control immobilization through site-specific labeling - to enable the development of high density arrays for proteomic applications. These improved multiplexed immunoassays will have significant value in both research and clinical diagnostics applications.

Thesaurus Terms:
Antibodies;Antibody Engineering;Antibody-Producing Cells;Avidin;Base;Binding (Molecular Function);Biological Assay;Biotinylation;Catalytic Domain;Cell Line;Cells;Chimera Organism;Chimeric Antibody;Chimeric Proteins;Clinical;Clinical Application;Clustered Regularly Interspaced Short Palindromic Repeats;Commercial Application;Confidential Information;Cost;Density;Design;Development;Diagnostic;Diagnostic Reagent;Evaluation;Fab Immunoglobulins;Gene Cloning;Generations;Genes;Genetic Recombination;Genome;Genomics;Goals;Government;Homologous Recombination;Hybridomas;Immobilization;Immunoassay;Immunoglobulin G;Immunoglobulin Genes;Improved;Label;Length;Link;Method Development;Methodology;Methods;Modification;Mus;New England;Nonhomologous Dna End Joining;O(6)-Methylguanine-Dna Methyltransferase;Oligonucleotide Microarrays;Oligonucleotides;Performance;Persons;Phase;Production;Programs;Property;Protein Array Analysis;Proteins;Proteomics;Protocols Documentation;Public Health Relevance;Rapid Technique;Reagent;Recombinant Antibody;Recombinants;Research;S-Nitro-N-Acetylpenicillamine;Self Assembly;Site;Small Business Innovation Research Grant;Streptavidin;System;Tertiary Protein Structure;Transfection;Validation;