定量放射性薄层色谱与正电子发射断层扫描测定链霉亲和素转导的 CAR-T 细胞的研究

The proliferation of chimeric antigen receptor (CAR) T cells is closely related to their efficacy, but it is still a great challenge to monitor and quantify CAR T cells in vivo. Based on the high affinity (Kd 10 -15 M) of streptavidin (SA) and biotin, radiolabeled biotin may be used to quantify SA-transduced CAR T cells (SA-CAR T cells). Radio-thin-layer chromatography (radio-TLC) and positron emission tomography (PET) are highly sensitive for trace analysis.

Our aim was to develop radio-TLC and PET methods to quantify SA-CAR T cells in vitro and in vivo. First, we developed [ 68 Ga]-DOTA-biotin. Commercially available SA was used as a standard, and quantitative standard curves were established in vitro and in vivo by radio-TLC and PET.

Furthermore, the feasibility of the method was verified in Raji model mice. The linear range of radio-TLC was 0. 02 0. 15 pmol/ L with R 2 = 0. 9993 in vitro. The linear range of PET was 0. 02 0. 76 pmol/ L with R 2 = 0. 9986 in vivo. SA in CAR T cells can also be accurately quantified in a Raji leukemia model according to PET imaging. The radio-TLC/PET method established in this study is promising for using in the dynamic monitoring and analysis of SA-CAR T cells during therapy.