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– CITP fractionation |
CITP Peptide Focusing Gemini™ leverages a novel electrokinetic focusing technology that takes advantage of transient capillary isotachophoresis/capillary zone electrophoresis (CITP/CZE) to perform complex proteome analysis for clinical relevant biological samples. Capillary zone electrophoresis (CZE) resolves proteins and peptides based on their differences in electrophoretic mobility which is a function of the charge-to-size ratio. Transient capillary isotachophoresis (CITP) is employed for on-capillary preconcentration of peptides prior to CZE separation. CITP, unlike most separation modes in capillary electrophoresis, employs a discontinuous electrolyte system. The stacking and separation of peptides and proteins occur in the boundary region between the leading electrolyte (LE) and terminating electrolytes (TE) under the influence of an applied electric field. The combination of sample concentration/preparation in-line with the subsequent electrophoretic separations avoids sample loss or dilution due to manual handling and transfer procedures. The unique characteristic inherent to CITP/CZE enables the successful study of clinical specimens with small sample volume to obtain a high-resolution analysis. Gemini™'s CITP/CZE separations significantly benefit the analysis of low abundance peptides by enhancing the dynamic range and detection sensitivity of MS. Combined with its high throughput and excellent resolving power in an automated platform, the coupling of capillary electrophoresis with nanoLC-MS used in the Gemini™ platform offers unique benefits toward effective global proteomic analysis through selective enrichment of peptides. Selective enrichment during CITP/CZE focusing Unlike MuDPIT separations using SCX, Gemini™'s CITP dimension allows the analysis of multiple-charged molecules, and prevents the overlap of peptides between multiple fractions, thereby ensuring that peptides from one fraction do not connvolute the analysis from other fractions. As a result, typical Gemini&trade analyses result in 3,000 &ndash 4,000 protein IDs at a stringent 1% false positive rate. Together with proprietary sample processing methods, CITP focusing also renders Gemini™ compatible with minute sample amounts. Key benefits of Gemini&trade's CITP technology:
Comparison of CITP and SCX separations
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