Publication details
Splitless gradient nanocolumn liquid chromatographic system for proteomic purposes
| Authors | |
|---|---|
| Year of publication | 2012 |
| Type | Conference abstract |
| Citation | DUŠA, Filip, Jozef ŠESTÁK, Dana MORAVCOVÁ, Marie HORKÁ and Vladislav KAHLE. Splitless gradient nanocolumn liquid chromatographic system for proteomic purposes. In 36th ISCC 2012, Riva del Garda, Italy, Abstract Book. 2012. |
| Description | Splitless and reproducible preparation of mobile phase gradients for nanocolumn liquid chromatography at the flow rates of 1 uL/min and less is not an easy task. Commercial systems could be used; nevertheless, these are bulky and expensive. Recently we have published a simple laboratory set-up for proteomic purposes for peptide pre-concentration and separation by gradient elution. It is based on the syringe infusion pump equipped with a 100-uL glass syringe in which the mobile phase gradient is prepared by successive sucking strong and weak mobile phases. In the present contribution we describe a fully automated splitless gradient nanocolumn liquid chromatographic system based on principle mentioned above. It consists of syringe pump and ten-port selector valve controlled by a computer. The Labview software is used to control all functions of the designed system which is capable to create a splitless gradient, to inject large sample volumes for sample pre-concentration, and quick fluid interchange with extremely low delay volume is possible. In the field of proteomics the separation techniques are essential for the most complete identification of proteins. We performed experiments with proteins extracted from bacterial cell surface. System proved that it has high efficiency and flexibility in analysis of whole intact proteins as well as in analysis of tryptic peptides. Consequently separated peptides were analyzed with MS and identified after database comparison. |