Highly efficient peptide separations in proteomics

Tryptic digest analysis by comprehensive reversed phase×two reversed phase liquid chromatography (RP‐LC×2RP‐LC) at different pH's

“…In a series of reports, we described a highly efficient 1D-LC system consisting of a series of conventional columns operated at elevated temperature [76][77][78] for the analysis of complex protein samples. One particular essay described the extension of the COFRA-DIC procedure (COmbined FRActional DIagonal Chromatography) to a column length of 1 m [76].…”

Performance evaluation of long monolithic silica capillary columns in gradient liquid chromatography using peptide mixtures

“…Since there is a high demand for an increased separation performance in the field of proteomics [7,8], monolithic silica capillary columns have also been explored for the separation and identification of complex peptide mixtures. While early reports focused on shorter columns [9,10], more recent papers deal with the use of long monolithic columns to further enhance the chromatographic performance and increase the number of identified peptides/proteins [11,12].…”

Recent advances on multidimensional liquid chromatography–mass spectrometry for proteomics: From qualitative to quantitative analysis—A review

“…A typical solution is to increase the column length or decrease the particle size. Although column technologies have seen major breakthroughs recently [8][9][10][11], a better and more straightforward method is to develop multidimensional HPLC (MDLC), by which proteins or peptides are separated by two or more independent separation mechanisms in a consecutive manner. The theoretical peak capacity (P c ) of a completely orthogonal MDLC system is defined as the product of the individual peak capacity from each dimension: P c1 × P c2 × .…”

“…Some excellent reviews summarized the development of MDLC systems and their applications in proteome analysis [10,[35][36][37][38][39][40][41][42][43][44][45][46][47], most of which focused on the discussion on maximizing the overall peak capacity of separation or applications of MDLC systems in proteomic studies. However, the predominant bottlenecks of modern MDLC platforms have changed from limited peak capacity to low detection limit and analysis throughput for the in-depth qualitative analysis, as well as the poor accuracy and precision of quantitative analysis of complex proteome samples.…”