This review article is an update of our previous one by C. Puangpila, E. Mayadunne, and Z. El Rassi, Electrophoresis 2015, 36, 238-252. Similarly to the previous article, this review has two main topics, including (i) proteomic sample preparation (e.g., depletion of high-abundance proteins, reduction of the protein dynamic concentration range, and enrichment of a particular subproteome), and (ii) the subsequent chromatographic and/or electrophoretic prefractionation prior to protein separation and identification by LC-MS/MS. More than 70 papers published in the period extending from mid-2014 to the present have been reviewed. Although an effort was made to yield a comprehensive review article, one may safely state that this review article is by no means thorough, and the aim was to rather provide a concise description of the latest developments in the field.