Current therapeutic antibodies target the cell surface and within the external cellular environment. Yet intracellular proteins make up half of the proteome. Having a facile means of targeting those intracellular proteins for the generation of therapeutic antibodies would be highly desirable. Targeting MHC complexed with neoantigens from intracellular proteins would greatly allow the generation of T cell receptor mimic (TCRm) antibodies for development as therapeutic drugs. We have developed a unique immune-targeting strategy that we call âEpivolveâ that can be used to make âsite directedâ Abs by leveraging the modularity of the complementarity determining region (CDR)-H2 of an Ab. Epivolve functions through the âevolutionâ of an Ab paratope using methods that are described in the proposal. The advantages of the in vivo method proposed include the isolation of Abs that have a high affinity (in the low picomolar range) for the peptide:MHC complex. This is because Abs can undergo multiple Ag challenges and affinity maturation via somatic hyper-mutagenesis in vivo to generate antibodies with single digit pM to hundredth nM. In vivo methods can also take advantage of transgenic animals for deriving human Abs. Affinities of TCRm Abs produced through phage display tend to lie in the moderate nanomolar range (â50â300 nM) and require further in vitro protein engineering. For this proposal, Abbratech will apply its novel site-directed Epivolve technology along with an improved rabbit single B cell sorting platform for the efficient discovery of TCRm Abs. The Epivolve method overcomes tolerance. And one of our goals is to generate antibodies in as little as 5 weeks. High-affinity, neoantigen:MHC-specific TCRm Abs have proven difficult to produce in large numbers by either traditional phage or hybridoma approaches. Enhanced technologies for the generation of TCRm Abs within a short period of time offer exciting opportunities to accelerate future TCRm Ab discovery.
Public Health Relevance Statement: NARRATIVE Current therapeutic antibodies target the cell surface and proteins within the external cellular compartments. Yet intracellular proteins make up half of the proteome. Having a facile means of targeting those intracellular proteins for the generation of therapeutic antibodies would be highly desirable. We have developed a unique immune-targeting strategy that we call âEpivolveâ that can be used to make âsite directedâ Abs. For this proposal, Abbratech will apply its novel Epivolve technology along with an improved rabbit single B cell sorting platform for the efficient discovery of potential therapeutic antibody drugs. The Epivolve method overcomes tolerance. And one of our goals is to generate antibodies in as little as 5 weeks. High-affinity Abs targeting intracellular regions of proteins placed within complexes on the cell surface are known as T cell receptor mimics (TCRms). TCRms have proven difficult to produce in large numbers by either traditional phage or hybridoma approaches. Enhanced technologies to generate TCRms within a short period of time offer exciting opportunities to accelerate future therapeutic discoveries.
Project Terms: Acceleration; Alanine; Allelomorphs; Alleles; Animals; Transgenic Animals; Antibodies; immunogen; Antigens; B-Lymphocytes; B blood cells; B cell; B cells; B-Cells; B-cell; Bacteriophages; Phages; bacterial virus; Antibody Binding Sites; Paratopes; antibody combining site; Blood; Blood Reticuloendothelial System; Cell Separation; Cell Isolation; Cell Segregation; Cell Separation Technology; cell sorting; Cells; Cell Body; Cohort Studies; Concurrent Studies; Discrimination; Cognitive Discrimination; Disease; Disorder; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Engineering; Environment; Euthanasia; Mercy Killing; Evolution; Exhibits; Future; Genes; Goals; Half-Life; Health; Hemorrhage; Bleeding; blood loss; Human; Modern Man; Hybridomas; Immunoglobulin G; 7S Gamma Globulin; IgG; Immunization; Complementarity Determining Regions; Complimentarity Determining Region; Hypervariable Loop; Hypervariable Regions; Immunoglobulin Hypervariable Region; In Vitro; Kinetics; Maps; Methods; Mus; Mice; Mice Mammals; Murine; United States National Institutes of Health; NIH; National Institutes of Health; Peptides; inorganic phosphate; Phosphates; Phosphopeptides; Pilot Projects; pilot study; pressure; Protein Engineering; genetic protein engineering; protein design; Proteins; Publishing; Oryctolagus cuniculus; Domestic Rabbit; Rabbits; Rabbits Mammals; Reagent; T-Cell Receptor; MHC Receptor; Major Histocompatibility Complex Receptor; T-Cell Antigen Receptors; Role; social role; Solvents; Specificity; Spleen; Spleen Reticuloendothelial System; Technology; Testing; Time; Generations; density; improved; Site; Biochemical; Measurement; Therapeutic; Nature; Complex; Scanning; Digit structure; Digit; hydrophilicity; cohort; MHC antigen; Peripheral Blood Mononuclear Cell; PBMC; novel; Cell surface; Proteome; Sampling; Property; cross reactivity; immunogenic; Enhancement Technology; HLA-A*0201; HLA A*0201 antigen; Molecular Interaction; Binding; Normal Cell; HLA-A gene; HLA-A; HLAA; Peptide/MHC Complex; Peptide-MHC; Peptide-Major Histocompatibility Protein Complex; pMHC; Affinity; Immune Targeting; Protein Region; in vivo; Collection; Phage Display; Tumor Biology; Monitor; Process; sex; Authorization documentation; Authorization; Permission; Development; developmental; Population; Therapeutic antibodies; tumor; overexpress; overexpression; nano-molar; nanomolar; experiment; experimental research; experiments; experimental study; Immunize; neo-antigen; neo-epitopes; n
