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.
22-mar-2022
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore CHIM/01 - CHIMICA ANALITICA
English
Con Impact Factor ISI
This work was in part supported by Associazione Italiana per la Ricerca sul Cancro, AIRC (project n. 14420) (F.R.), by the European Research Council, ERC (Consolidator Grant project n. 819160) (F.R.), by the Marie Skłodowska-Curie grant agreement (“DNA-bots” project n. 843998) (T.P.), by the Marie Skłodowska-Curie grant agreement (“DNA-NANO-AB” project n. 843179) (S.R.) and by the Marie Skłodowska-Curie ITN project DNA-robotics (“DNA-robotics”, project n. 765703) (F.R. and A.P.D.)
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].
Patino Diaz, A; Bracaglia, S; Ranallo, S; Patino, T; Porchetta, A; Ricci, F
Articolo su rivista
File in questo prodotto:
File Dimensione Formato  
Programmable_transcriptional_TEXT.pdf

accesso aperto

Tipologia: Documento in Pre-print
Licenza: Non specificato
Dimensione 968.2 kB
Formato Adobe PDF
968.2 kB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/299516
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 26
  • ???jsp.display-item.citation.isi??? 25
social impact