An Automated High-Throughput Sample Preparation Protocol for LC-MS/MS Analysis of Glycopeptides

Title:An Automated High-Throughput Sample Preparation Protocol for LC-MS/MS Analysis of Glycopeptides

Volume: 13 Issue: 1

Author(s): Albert-Baskar Arul, Jong-Moon Park, Hookeun Lee, Jeong-Heum Baek, Jongho Jeon, Eunhee Ji, Jae Won Oh and Kwang Pyo Kim

Affiliation:

Keywords: FACE, FASP, glycoproteome, high-throughput, Lectin enrichment, membrane filter plate, N-glycopeptide, sample preparation.

Abstract: Current sample preparation workflows for glycopeptide analyses have low levels of automation and low sample throughput. In this study, we have developed an automated glycopeptide preparation workflow for a 96-well plate liquid handling robotic system. The protocol is based on the filter-aided capture and elution method. The glycopeptides bound to lectins were trapped on the filter, and the filters were then incubated with PNGaseF to release the glycopeptides specifically. The experimental conditions were optimized for lectin: peptide ratios using a standard glycoprotein and colon tissue proteomes. Duplicates of ten samples were processed in parallel and subjected to LC-MS/MS analysis. The results of LC-MS/MS analysis showed that 96 samples can be processed in parallel in 6.5 hours.

Cite this article as:

Arul Albert-Baskar, Park Jong-Moon, Lee Hookeun, Baek Jeong-Heum, Jeon Jongho, Ji Eunhee, Oh Won Jae and Kim Pyo Kwang, An Automated High-Throughput Sample Preparation Protocol for LC-MS/MS Analysis of Glycopeptides, Current Proteomics 2016; 13 (1) . https://dx.doi.org/10.2174/1570164613666160413125317