Construction and characterisation of a functional CD19 specific single chain Fv fragment for immunotherapy of B lineage leukaemia and lymphoma

Mol Immunol. Nov-Dec 1997;34(16-17):1157-65. doi: 10.1016/s0161-5890(97)00144-2.

Abstract

The B cell specific antigen CD19 is a target for the immunotherapy of B lineage leukaemias and lymphomas. We have engineered a single chain Fv (scFv) fragment from the mouse hybridoma cell line FMC63 which produces monoclonal antibody specific for CD19. The genes encoding the FMC63 heavy and light chain variable regions were amplified from cDNA and a scFv was constructed by splice overlap extension PCR. Analysis of staining of lymphoblastoid cell lines, peripheral blood lymphocytes and tonsil sections demonstrated that the monovalent scFv fragment has the same cellular specificity as the parent hybridoma antibody. Kinetic studies with radiolabelled material showed that the scFv binds target cells with a Ka of 2.3 x 10(-9), compared with 4.2 x 10(-9) for the parent antibody. This CD19 scFv will be used in experimental models to test its therapeutic efficacy and immunogenicity, with a view to application in the diagnosis and treatment of human B cell cancers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD19 / immunology*
  • Base Sequence
  • DNA, Complementary / genetics
  • DNA, Complementary / immunology
  • Humans
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / immunology*
  • Immunoglobulin Fragments / therapeutic use
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology
  • Immunotherapy*
  • Leukemia, B-Cell / immunology*
  • Leukemia, B-Cell / therapy
  • Lymphoma, B-Cell / immunology*
  • Lymphoma, B-Cell / therapy
  • Mice
  • Molecular Sequence Data
  • Protein Engineering

Substances

  • Antigens, CD19
  • DNA, Complementary
  • Immunoglobulin Fragments
  • Immunoglobulin Variable Region

Associated data

  • GENBANK/Y14283
  • GENBANK/Y14284