Elsevier

Molecular Immunology

Volume 34, Issues 16–17, November–December 1997, Pages 1157-1165
Molecular Immunology

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

https://doi.org/10.1016/S0161-5890(97)00144-2Get rights and content

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 × 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.

References (46)

  • F.M. Uckun et al.

    Detailed studies on expression and function of CD19 surface determinant by using B43 monoclonal antibody and the clinical potential of anti-CD19 immunotoxins

    Blood

    (1988)
  • P. Antoniw et al.

    Radioimmunotherapy of colorectal carcinoma xenografts in nude mice with yttrium-90 A33 IgG and Tri-Fab (TFM)

    Br. J. Cancer

    (1996)
  • C. Bebbington

    Expression of antibody genes in mammalian cells

  • R.H. Begent et al.

    Clinical evidence of efficient tumor targeting based on single-chain Fv antibody selected from a combinatorial library

    Nat. Med.

    (1996)
  • B.E. Bejcek et al.

    Development and characterization of three recombinant single chain antibody fragments (scFvs) directed against the CD19 antigen

    Cancer Res.

    (1995)
  • A.E. Bolton et al.

    The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent

    Biochem. J.

    (1973)
  • P. Carter et al.

    Improved oligonucleotide site-directed mutagenesis using M13 vectors

    Nucleic Acids Res.

    (1985)
  • J.L. Casey et al.

    Preparation, characterisation and tumour targeting of cross-linked divalent and trivalent anti-tumour Fab′ fragments

    Br. J. Cancer

    (1996)
  • T. Clackson et al.

    General applications of PCR to gene cloning and manipulation

  • B. Dorken et al.

    B cell antigens: CD19

  • G.I. Evan et al.

    Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product

    Mol. Cell Biol.

    (1985)
  • J.V. Gavilondo-Cowley et al.

    Specific amplification of rearranged immunoglobulin variable region genes from mouse hybridoma cells

    Hybridoma

    (1990)
  • M.L. Grossbard et al.

    Anti-B4-blocked ricin: a phase I trial of 7-day continuous infusion in patients with B-cell neoplasms

    J. Clin. Oncol.

    (1993)

    • Cited by (0)

    §

    Present Address: Department of Development and Genetics, Babraham Institute, Babraham, Cambridge, CB2 4AT, U.K.

    View full text
    Copyright © 1997 Published by Elsevier Ltd.