Recent Advancements and Applications of Size Exclusion Chromatography in Modern Analysis

Abstract

Abstract: In recent years, size exclusion chromatography (SEC) has gained valuable and impactable recognition among various chromatographic techniques. Also addressed as other names, viz. gel permeation chromatography, steric-exclusion chromatography, etc., SEC is typically taken into consideration for the fractionation and molecular weight determination of biomolecules and large macromolecules (proteins and polymers) using porous particles. A homogenous mixture of molecules dispersed in the mobile phase is introduced to the chromatographic column, which provides a solid support in the form of microscopic beads (the stationary phase). The beads act as “sieves” and purify small molecules, which become temporarily trapped inside the pores. Some of the advantages that SEC offers over other chromatographic techniques are short analysis time, no sample loss, good sensitivity, and requirement for less amount of mobile phase. In the proposed manuscript, we have deliberated various proteomic applications of size exclusion chromatography, which include the isolation of extracellular vesicles in cancer, isolation of human synovial fluid, separation of monoclonal antibodies, as well as several tandem techniques, such as deep glycoproteomic analysis using SEC-LC-MS/MS, analysis of mammalian polysomes in cells and tissues using tandem MS-SEC, SEC-SWATH-MS profiling of the proteome with a focus on complexity, etc.