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A novel luciferase-based assay for the detection of Chimeric Antigen Receptors.

Sci Rep. 2019-02; 
GopalakrishnanRamakrishnan,MattaHittu,ChoiSunju,NatarajanVenkatesh,PrinsRuben,GongSongjie,ZenunovicArta,NarasappaNell,PatelFatima,PrakashRekha,ChaudharyVishan,SikriVarun,ChitnisSaurabh Deepak,KochegarovAndrei,WangDan,FalatMagdalena,KahnMichael,KeerthipatiPooja Smruthi,SharmaNaman,LenkaJyotirmayee,StiebenTomas Meza,BraunJason,BatraAnkita,PurvisKatelyn,ItoKenta,LeeJae Han,JeronimoAlberto,ZamoraHannalei Mae,MembrenoAllen,QiuQueenie,PeshinSupriya,NamburuLalith,ChaudharyPre
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摘要

Chimeric Antigen Receptor-T (CAR-T) cell immunotherapy has produced dramatic responses in hematologic malignancies. One of the challenges in the field is the lack of a simple assay for the detection of CARs on the surface of immune effector cells. In this study, we describe a novel luciferase-based assay, termed Topanga Assay, for the detection of CAR expression. The assay utilizes a recombinant fusion protein, called Topanga reagent, generated by joining the extra-cellular domain of a CAR-target in frame with one of the marine luciferases or their engineered derivatives. The assay involves incubation of CAR expressing cells with the Topanga reagent, a few washes and measurement of luminescence. The... More

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