High-Resolution Ultrahigh-Pressure Long Column Reversed-Phase Liquid Chromatography for Top-down Proteomics — SMT REFERENCED ARTICLE

Yufeng Shen, Nikola Tolić, Paul D. Piehowski, Anil K. Shukla, Sangtae Kim, Rui Zhao, Yi Qu, Errol Robinson, Richard D. Smith
Journal of Chromatography A

Journal-of-Chromatography-A Separation Methods TechnologiesSeparation of proteoforms for global intact protein analysis (i.e. top-down proteomics) has lagged well behind what is achievable for peptides in traditional bottom-up proteomic approach and is becoming a true bottle neck for top-down proteomics. Herein, we report use of long (≥1 M) columns containing short alkyl (C1-C4) bonded phases to achieve high-resolution RPLC for separation of proteoforms…The packing…silica particles, were from…Separation Methods Technologies (Newark, DE) [3µm, 300Å C1 porous silica (BMEB1-3-300) and 3µm, 300Å C2 porous silica (BMEB2-3-300)]…C1 RPLC has the greatest potential in terms of elution of hydrophobic and larger proteoforms; accordingly, we observed dense peak arrays in the C1 RPLC late elution zone…READ MORE HERE