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Development and Characterization of Three Recombinant Single Chain Antibody Fragments (scFvs) Directed against the CD19 Antigen¹

Bone Marrow Transplant Program and Departments of Pediatrics and Laboratory Medicine/Pathology [B. E. B., D. W., E. B., C. A. P., F. M. U., J. H. K.], and Department of Therapeutic Radiology [F. M. U.], University of Minnesota, Minneapolis, Minnesota 55455; Brown Cancer Center, Louisville, Kentucky 40292 [S. C. P.]; and Cross Cancer Institute, University of Alberta, Edmonton, Alberta, Canada [S. P.]

Antibodies that recognize antigens restricted to leukemia, lymphoma, and normal hematopoietic cells represent a unique opportunity to develop therapeutics, because they have the potential for relatively selective treatment of these diseases. Antibodies that recognize the CD19 antigen found on normal and malignant B cells, but not on stem cells, have been used to develop immunoconjugates. However, these conjugates are large and might be suboptimal in tumor penetration when compared to molecules using smaller single chain Fv (scFv) antibody fragments. scFv has the advantage of being a molecularly engineered homogeneous molecule. In this report, we demonstrate the cloning, expression, and binding of three anti-CD19 antibodies as scFvs. All three scFvs were successfully cloned and expressed. FVS191, derived from cell line B43, and FVS192, derived from SJ25C1, were properly refolded and bound CD19 antigen in FACS competition assays. These anti-CD19 scFv should be useful in the further development of diagnostic and therapeutic molecules.

¹ Supported in part by an Outstanding Investigator Grant from the National Cancer Institute (CA 49721; to J. H. K.). F. M. U. is a scholar of the Leukemia Society of America.

² To whom requests for reprints should be addressed, at Box 86 UMHC, 420 Delaware Street, SE, Minneapolis, MN 55455.

Received 10/17/94. Accepted 4/ 5/95.

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