Amelogenins are structural proteins of the dental enamel matrix, playing a key role in archaeological and forensic fields due to their relative abundance and relevance for sex determination. Currently, the detection of amelogenins mainly relies on liquid chromatography (LC) and tandem mass spectrometry (MS/MS) and only few studies have involved immunoassays based on antibodies, but the application of fast, reusable and cost-effective biomimetic systems has not yet been explored. We present an innovative biosensor based on Molecularly Imprinted Bio-Polymers (MIBPs) coupled with Surface Plasmon Resonance (SPR), capable of detecting and binding amelogenins from standard solutions and protein extracts from human teeth samples. The protein sequence was studied by machine learning methods to select suitable peptide sequences to be used to molecularly imprint polynorepinephrine (PNE). The imprinting efficiency of the PNE-based biosensor was evaluated by performing single cycle kinetic (SCK) analyses, allowing the characterisation of the system in terms of kinetic rates and binding affinity parameters, towards both the peptide template and protein. The demonstration of the applicability in real cases was achieved optimising a suitable extraction protocol and performing measurements of extracts from human dental enamel. The results prove the effectiveness of the biosensor, which was specifically tailored on the target protein sequence and requires a quick and simple preparation process involving natural, 100% green, and biodegradable reagents. The application of state-of-the-art MIBPs offers a low-cost, highly effective, and environmentally friendly alternative to the use of antibodies in the development of label-free detection systems that allows the possibility to select and design the epitope to be printed with high precision, therefore deciding the protein capture target.
Development of a new biomimetic biosensor based on Molecularly Imprinted Bio-Polymers coupled with Surface Plasmon Resonance for the detection of amelogenins / Camagni, Valentina; Sestaioni, Davide; Gatti, Lucrezia; Sciutto, Giorgia; Prati, Silvia; Mazzeo, Rocco; Scarano, Simona. - (2024). (Intervento presentato al convegno XXVIII Congresso Nazionale SCI2024 tenutosi a Milano).
Development of a new biomimetic biosensor based on Molecularly Imprinted Bio-Polymers coupled with Surface Plasmon Resonance for the detection of amelogenins
Valentina Camagni
Primo
;
2024
Abstract
Amelogenins are structural proteins of the dental enamel matrix, playing a key role in archaeological and forensic fields due to their relative abundance and relevance for sex determination. Currently, the detection of amelogenins mainly relies on liquid chromatography (LC) and tandem mass spectrometry (MS/MS) and only few studies have involved immunoassays based on antibodies, but the application of fast, reusable and cost-effective biomimetic systems has not yet been explored. We present an innovative biosensor based on Molecularly Imprinted Bio-Polymers (MIBPs) coupled with Surface Plasmon Resonance (SPR), capable of detecting and binding amelogenins from standard solutions and protein extracts from human teeth samples. The protein sequence was studied by machine learning methods to select suitable peptide sequences to be used to molecularly imprint polynorepinephrine (PNE). The imprinting efficiency of the PNE-based biosensor was evaluated by performing single cycle kinetic (SCK) analyses, allowing the characterisation of the system in terms of kinetic rates and binding affinity parameters, towards both the peptide template and protein. The demonstration of the applicability in real cases was achieved optimising a suitable extraction protocol and performing measurements of extracts from human dental enamel. The results prove the effectiveness of the biosensor, which was specifically tailored on the target protein sequence and requires a quick and simple preparation process involving natural, 100% green, and biodegradable reagents. The application of state-of-the-art MIBPs offers a low-cost, highly effective, and environmentally friendly alternative to the use of antibodies in the development of label-free detection systems that allows the possibility to select and design the epitope to be printed with high precision, therefore deciding the protein capture target.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
