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 Blood, 15 April 2007, Vol. 109, No. 8, pp. 3393-3399.
Prepublished online as a Blood First Edition Paper on December 12, 2006; DOI 10.1182/blood-2006-07-030593.

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IMMUNOBIOLOGY

Cell-free production of scFv fusion proteins: an efficient approach for personalized lymphoma vaccines

Gregory Kanter1, Junhao Yang2, Alexei Voloshin2, Shoshana Levy1, James R. Swartz2,3, and Ronald Levy1

1 Department of Medicine, Division of Oncology, Stanford University Medical Center, Stanford, CA; 2 Department of Chemical Engineering, Stanford University, Stanford, CA; 3 Department of Bioengineering, Stanford University, Stanford, CA

The unique immunoglobulin (Ig) idiotype on the surface of each B-cell lymphoma represents an ideal tumor-specific antigen for use as a therapeutic vaccine. We have used an Escherichia coli—based, cell-free protein-expression system to produce a vaccine within hours of cloning the Ig genes from a B-cell tumor. We demonstrated that a fusion protein consisting of an idiotypic single chain Fv antibody fragment (scFv) linked to a cytokine (GM-CSF) or to an immunostimulatory peptide was an effective lymphoma vaccine. These vaccines elicited humoral immune responses against the native Ig protein displayed on the surface of a tumor and protected mice against tumor challenge with efficacy equal to that of the conventional Ig produced in a mammalian cell and chemically coupled to keyhole limpet hemocyanin. The cell-free E coli system offers a platform for rapidly generating individualized vaccines, thereby allowing much more efficient application in the clinic.


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