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Bispecific CD3 × CD19 diabody for T cell‐mediated lysis of malignant human B cells
Author(s) -
Kipriyanov Sergey M.,
Moldenhauer Gerhard,
Strauss Gudrun,
Little Melvyn
Publication year - 1998
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19980831)77:5<763::aid-ijc16>3.0.co;2-2
Subject(s) - microbiology and biotechnology , cd19 , chemistry , cd3 , antibody , antigen , immunotoxin , lysis , flow cytometry , cytotoxicity , periplasmic space , biology , escherichia coli , biochemistry , in vitro , immunology , cd8 , gene
For the treatment of minimal residual disease in patients with leukemias and malignant lymphomas, we constructed a heterodimeric diabody specific for human CD19 on B cells and CD3ϵ chain of the T cell receptor complex. The bispecific diabody was expressed in Escherichia coli using a vector containing a dicistronic operon for co‐secretion of V H 3‐V L 19 and V H 19‐V L 3 single‐chain Fv fragments (scFv). It was purified in one step by immobilized metal affinity chromatography (IMAC) from the periplasmic extract and culture medium. Flow cytometry experiments revealed specific interactions of the diabody with both CD3 and CD19 positive cells, to which it bound with affinities close to those of the parental scFvs. It was less stable than anti‐CD3 scFv but more stable than anti‐CD19 scFv when incubated in human serum at 37°C. In cytotoxicity tests, the diabody proved to be a potent agent for retargeting peripheral blood lymphocytes to lyse tumor cells expressing the CD19 antigen. The efficiency of cell lysis compared favorably with that obtained with a bispecific antibody (BsAb) of the same dual specificity that was prepared by the quadroma technique. Int. J. Cancer 77:763–772, 1998. © 1998 Wiley‐Liss, Inc.