Multilayer gold nanoparticle-assisted protein tryptic digestion in solution and in gel under photothermal heating

Elevating the reaction temperature is an effective method to accelerate protein enzymatic digestion because it can promote protein denaturation and enzyme activities. In this study, we demonstrated a new photothermal heating method to assist protein tryptic digestion on glass slides. A glass slide coated with layer-by-layer gold nanoparticles (Glass@AuNPs), combined with the use of a near infrared (NIR) diode laser, was used to raise reaction temperature during tryptic digestion in a short period of time. The modified glass slide is capable of absorbing NIR light arising from the dipole-dipole interactions between Au NPs immobilized on the slide. The temperature of Glass@AuNPs rapidly increased when irradiated by the NIR laser, accelerating protein enzymatic digestion conducted on the slide. Thus, when performing the tryptic digestion of proteins on the Glass@AuNPs slide under NIR irradiation, 3.5 min was sufficient to carry out the tryptic digestion of proteins in solution, while less than 5 min was adequate for in-gel tryptic digestion of proteins. Matrix-assisted laser desorption/ionization mass spectrometry was used for characterization of the tryptic digestion product. On the basis of the results, the time taken to analyze proteins could be greatly reduced using this current approach.