Chimeric antigen receptor preparation from hybridoma to T-cell expression

Hakan Köksal; Elizabeth Baken; David John Warren; Geir Åge Løset; Else Marit Inderberg; Sébastien Wälchli

doi:10.1093/abt/tbz007

Chimeric antigen receptor preparation from hybridoma to T-cell expression

Antib Ther. 2019 May 31;2(2):56-63. doi: 10.1093/abt/tbz007. eCollection 2019 Apr.

Authors

Hakan Köksal¹, Elizabeth Baken¹, David John Warren², Geir Åge Løset^{3

4}, Else Marit Inderberg¹, Sébastien Wälchli¹

Affiliations

¹ Department of Cellular Therapy, Department of Oncology, Oslo University Hospital-Radiumhospitalet, Oslo 0379, Norway.
² Department of Medical Biochemistry, Oslo University Hospital-Radiumhospitalet, Oslo 0379, Norway.
³ Department of Biosciences, University of Oslo, Oslo 0316, Norway.
⁴ Nextera AS, Oslo, Norway.

Abstract

The successful use of chimeric antigen receptor (CAR) for hematological cancer treatment has influenced the direction taken in translational research toward an increasing focus on personalized targeted immunotherapy. Thus, a growing number of labs worldwide are now interested in testing their old antibody collections in this format to broaden the spectrum of utility and improve safety and efficacy. We herein present a straightforward protocol for the identification of an antibody from a hybridoma and the design of the single chain fragment that will be placed on the extracellular part of the CAR construct. We further show how to test the expression and the activity of the construct in primary T cells. We illustrate our demonstration with two new CARs targeted against the B cell receptor, more precisely the light chains κ and λ, that represent potential alternatives to the CD19 CAR used in the treatment of B-cell malignancies.

Keywords: antibody; chimeric antigen receptor; hybridoma; single chain variable fragment.