No abstract

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 assay can detect CARs comprising either immunoglobulin- or non-immunoglobulin-based antigen binding domains. We further demonstrate that addition of epitope tags to the Topanga reagent not only allows its convenient one step purification but also extends its use for detection of CAR cells using flow cytometry. However, crude supernatant containing the secreted Topanga reagent can be directly used in both luminescence and flow-cytometry based assays without prior protein purification. Our results demonstrate that the Topanga assay is a highly sensitive, specific, convenient, economical and versatile assay for the detection of CARs.

We examined the effect of two greenhouse covering materials (glass or solid polycarbonate sheets) on the light environment and growth of tomato and cucumber plants. Spectral analysis showed that polycarbonate sheets entirely blocked radiation in both the UV-B (300-320 nm) and UV-A (320-400 nm) ranges, whereas glass transmitted UV-A and was only opaque to UV-B. The transmittance of photosynthetically active radiation (400-700 nm) and near infrared radiation (700-1100 nm) was higher in polycarbonate than glass. Air and soil temperatures were not significantly different between greenhouses covered with either material. The growth of cucumber plants was slightly affected by covering materials, whereas no significant changes in growth parameters were observed for tomato plants. The color parameters of tomato fruits were affected by the cover material, whereas cucumber fruits showed similar coloration in both glass and polycarbonate greenhouses.

No abstract

No abstract