Molecularly imprinted polynorepinephrine for an innovative and sustainable approach for archaeological enamel analysis

Amelogenins (AMG) are crucial for sexing archaeological human remains, yet standard Mass Spectrometry analysis is often complex and costly. To address these issues, we present an innovative approach to imprint mimetic receptors based on Molecularly Imprinted Polynorepinephrine (PNE) able to detect and bind full-length AMG from standard solutions and AMG residues from enamel extracts. After establishing a simple and effective digestion-free protein extraction protocol from 1 to 2 mg of enamel powder samples, an original "Finger-Imprint" (FI) approach was attempted by directly imprinting the whole protein extract from modern and archaeological human samples. The biomimetic receptor was coupled with Surface Plasmon Resonance and Bio-Layer Interferometry platforms to assess imprinting efficiency, kinetic rates and affinity parameters of the system towards target proteins from standard solutions. NanoLC-MS/MS, μRaman and µFT-IR Spectroscopy were used to validate the extraction protocol, identifying the extract components and ensuring the absence of hydroxylapatite. The successful results prove the effectiveness of the FI-based biomimetic receptor, which was specifically tailored to the extract and requires a quick and streamlined preparation involving biocompatible and biodegradable reagents.