Optimization of Novel GELFrEE Fractionation for Molecular Weight–Based In-solution Protein Separation from Buffalo Meat, Pork, and Chicken

Peptidomics Approaches for Authentication of Pork and Water Buffalo Meat Employing Rapid In-solution Fractionation and 2-DE

“…The ability to load a relatively large volume of sample (max. 5.0 mg/strip) in GELFrEE yields impressive proteome coverage and is especially valuable during the analysis of low-abundance proteins, which have extremely important functions in various biological processes (Maheswarappa et al 2021; Banerjee et al 2021).…”

“…These researchers have demonstrated detection of as low as 1.0% w/w contaminating chicken in a pork meat mix using OFFGEL electrophoresis and detection of chicken-speci c peptides generated from myosin light chain 3 (MLC-3) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). However, OFFGEL electrophoresis requires a 24-36 h run time and the protein samples need to be puri ed prior to loading in order to remove the salts that may offer resistance while fractionating (Maheswarappa et al 2021;Sentandreu et al 2010) The GELFrEE fractionation has been reported by a group of researchers mainly for the separation and characterization of microbial proteins and biological samples (Melani et al 2016;Catherman et al 2013;Orton et al 2013;Kellie et al 2010;Doucette 2008, 2009). However, the GELFrEE approach has been minimally explored in meat/food protein fractionation, and recently our laboratory has successfully employed GELFrEE as a quick and rapid pre-fractionation tool prior to MS characterization of total and myo brillar/sarcoplasmic proteins from water buffalo meat, chicken and pork under raw and heatprocessed conditions (Maheswarappa et al 2021).…”

“…However, OFFGEL electrophoresis requires a 24-36 h run time and the protein samples need to be puri ed prior to loading in order to remove the salts that may offer resistance while fractionating (Maheswarappa et al 2021;Sentandreu et al 2010) The GELFrEE fractionation has been reported by a group of researchers mainly for the separation and characterization of microbial proteins and biological samples (Melani et al 2016;Catherman et al 2013;Orton et al 2013;Kellie et al 2010;Doucette 2008, 2009). However, the GELFrEE approach has been minimally explored in meat/food protein fractionation, and recently our laboratory has successfully employed GELFrEE as a quick and rapid pre-fractionation tool prior to MS characterization of total and myo brillar/sarcoplasmic proteins from water buffalo meat, chicken and pork under raw and heatprocessed conditions (Maheswarappa et al 2021). Our earlier study has encouraged us to explore the potential of GELFrEE for species authentication in complex biological samples like raw and heat-processed meat, and binary meat mixes.…”