We report here the development of a cell-free in-vitro transcription system for the detection of specific target antibodies. The approach is based on the use of programmable antigen-conjugated DNA-based conformational switches that, upon binding to a target antibody, can trigger the cell-free transcription of a light-up fluorescence-activating RNA aptamer. The system couples the unique programmability and responsiveness of DNA-based systems with the specificity and sensitivity offered by invitro genetic circuitries and commercially available transcription kits. We demonstrate that cell-free transcriptional switches can efficiently measure antibody levels directly in blood serum. Thanks to the programmable nature of the sensing platform the method can be adapted to different antibodies: we demonstrate here the sensitive, rapid and cost-effective detection of three different antibodies and the possible use of this approach for the simultaneous detection of two antibodies in the same solution.
Patino Diaz, A., Bracaglia, S., Ranallo, S., Patino, T., Porchetta, A., Ricci, F. (2022). Programmable cell-free transcriptional switches for antibody detection. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 144(13), 5820-5826 [10.1021/jacs.1c11706].
Programmable cell-free transcriptional switches for antibody detection
Ranallo, Simona;Porchetta, Alessandro;Ricci, Francesco
2022-03-22
Abstract
We report here the development of a cell-free in-vitro transcription system for the detection of specific target antibodies. The approach is based on the use of programmable antigen-conjugated DNA-based conformational switches that, upon binding to a target antibody, can trigger the cell-free transcription of a light-up fluorescence-activating RNA aptamer. The system couples the unique programmability and responsiveness of DNA-based systems with the specificity and sensitivity offered by invitro genetic circuitries and commercially available transcription kits. We demonstrate that cell-free transcriptional switches can efficiently measure antibody levels directly in blood serum. Thanks to the programmable nature of the sensing platform the method can be adapted to different antibodies: we demonstrate here the sensitive, rapid and cost-effective detection of three different antibodies and the possible use of this approach for the simultaneous detection of two antibodies in the same solution.File | Dimensione | Formato | |
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