Hyphenation of 2D heart cutting CIEF/CE-MS

SYDES, DANIEL; KLER, PABLO A.; LUTZ, D.; ZIPFL, P.; HUHN, CAROLIN

Congreso; CE Pharm 2015: CE in the Biotechnology & Pharmaceutical Industries: 17th Symposium on the Practical Applications for the Analysis of Proteins, Nucleotides and Small Molecules.; 2015

The hyphenation of capillary isoelectric focusing (CIEF) with mass spectrometry (MS) is a very promising tool regarding the characterization of proteins and peptides especially for R&D and quality control of monoclonal antibodies including their glycosylation. However, the coupling of CIEF with MS is still challenging due to the high number of carrier ampholytes (CAs) being incompatible with MS detection. Inthis work we seek to establish a 2D separation approach via CIEF/CE-MS toseparate analytes from these carrier ampholytes in the second CE separation column. A hybrid capillary-microchip-capillary module is used to connect the first and second dimension. Here we show the technical implementation, spacers and intermediate optical detection to control the positioning of the pH gradient in the interface within the 2D setup as well as first CIEF/CE-MS experiments.