Protein Downstream Processing

Calvano CD, De Ceglie C, Zambonin CG, Cosima Damiana Calvano, Cristina De Ceglie, Carlo G. Zambonin, Calvano, Cosima Damiana, Ceglie, Cristina De, Zambonin, Carlo G.

MALDI MS has become a technique of considerable impact on many fields, from proteomics to lipidomics, including polymer analysis and, more recently, even low molecular weight analytes due to the introduction of matrix-less ionization techniques (e.g., DIOS) or new matrices such as ionic liquids, proton sponges, and metal nanoparticles. However, protein identification by peptide mass fingerprint (PMF) still remains the main routine application. In the last few years, MALDI MS has played an emerging role in food chemistry especially in detection of food adulterations, characterization of food allergens, and investigation of protein structural modifications, induced by various industrial processes that could be detrimental for food quality and safety. Sample handling and pretreatment can be very different depending on the physical state, liquid or solid, of the analyzed matrices. Here, we describe simple protocols for protein extraction and MALDI MS analysis of liquid (milk) and solid (hazelnuts) samples taken as model. A classic approach based on a preliminary SDS gel electrophoresis separation followed by in-gel digestion and a faster approach based on in-solution digestion of whole samples are described and compared.