title: Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines.
abstract: The unique immunoglobulin (Ig) idiotype on the surface of each B-cell lymphoma represents an ideal tumor-specific antigen for use as a therapeutic vaccine. We have used an Escherichia coli-based, cell-free protein-expression system to produce a vaccine within hours of cloning the Ig genes from a B-cell tumor. We demonstrated that a fusion protein consisting of an idiotypic single chain Fv antibody fragment (scFv) linked to a cytokine (GM-CSF) or to an immunostimulatory peptide was an effective lymphoma vaccine. These vaccines elicited humoral immune responses against the native Ig protein displayed on the surface of a tumor and protected mice against tumor challenge with efficacy equal to that of the conventional Ig produced in a mammalian cell and chemically coupled to keyhole limpet hemocyanin. The cell-free E coli system offers a platform for rapidly generating individualized vaccines, thereby allowing much more efficient application in the clinic.
authors.affiliation: Department of Medicine, Division of Oncology, Stanford University Medical Center, Stanford, CA 94305, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Kanter
authors.forename: Junhao
authors.initials: J
authors.lastname: Yang
authors.forename: Alexei
authors.initials: A
authors.lastname: Voloshin
authors.forename: Shoshana
authors.initials: S
authors.lastname: Levy
authors.forename: James R
authors.initials: JR
authors.lastname: Swartz
authors.forename: Ronald
authors.initials: R
authors.lastname: Levy
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000528:radiation effects; D019496:Cancer Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D051379:Mice; D009374:Neoplasms, Experimental / Q000235:genetics / Q000276:immunology / Q000628:therapy; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D007130:Immunoglobulin Idiotypes; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor