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Effective lysis of lymphoma cells with a stabilised bispecific single‐chain Fv antibody against CD19 and Fc γ RIII (CD16)
Author(s) -
Bruenke Joerg,
Barbin Karin,
Kunert Susanne,
Lang Peter,
Pfeiffer Matthias,
Stieglmaier Kristin,
Niethammer Dietrich,
Stockmeyer Bernhard,
Peipp Matthias,
Repp Roland,
Valerius Thomas,
Fey Georg H.
Publication year - 2005
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.2005.05414.x
Subject(s) - cd19 , cd16 , antibody , antigen , b cell , microbiology and biotechnology , fc receptor , immunology , biology , chemistry , cd3 , cd8
Summary A recombinant bispecific single‐chain fragment variable antibody (bsscFv), directed against the B‐cell antigen CD19 and the low affinity Fc‐receptor Fc γ RIII (CD16), was designed for use in the treatment of patients with leukaemias and lymphomas. The Fc‐portions of whole antibodies were deliberately eliminated in this construct to avoid undesired effector functions. A stabilised bsscFv, ds[CD19 × CD16], was generated, in which disulphide bonds bridging the respective variable light (VL) and variable heavy (VH) chains were introduced into both component single‐chain (sc)Fvs. After production in 293T cells and chromatographic purification, ds[CD19 × CD16] specifically and simultaneously bound both antigens. The serum stability of ds[CD19 × CD16] was increased more than threefold when compared with the unstabilised counterpart, while other biological properties were not affected by these mutations. In antibody‐dependent cellular cytotoxicity experiments, ds[CD19 × CD16] mediated specific lysis of both CD19‐positive malignant human B‐lymphoid cell lines and primary tumour cells from patients with B‐cell chronic lymphocytic leukaemia or B‐cell acute lymphoblastic leukaemia. Natural killer cells, mononuclear cells (MNCs) from healthy donors and, in some instances, MNCs isolated from patients after allogeneic stem cell transplantation, were used as effectors. Thus, ds[CD19 × CD16] holds promise for the treatment of CD19 + B‐lineage malignancies.