On-Line Dual Microdialysis with ESI-MS for Direct Analysis of Complex Biological Samples and Microorganism Lysates

A novel dual-microdialysis approach has been developed for fast and efficient fractionation and cleanup for ESI-MS and ESI MS/MS analyses of biological samples. A modified dynamic microdialyzer utilizing two mirror-image serpentine channels, which sandwich a regenerated cellulose membrane of selected molecular weight cutoff, serves as the first stage for the removal of high-molecular-weight components and cellular residue. The second stage employs a hollow microdialysis capillary to remove low-molecular-weight species (e.g., salts) which can degrade or preclude analysis ESI-MS. A protein mixture consisting of 30 microM bovine serum albumin (BSA), 4.0 microM cytochrome c, 2.3 microM ubiquitin, and 9.4 microM bradykinin in 0.5 M NaCl was used to evaluate the performance of this system. Essentially complete removal of both BSA and NaCl was achieved, resulting in high-quality mass spectra containing only the lower molecular weight proteins. After passing through the on-line dual-microdialysis system, a crude bacteria cell lysate yielded clean ESI-mass spectra in approximately 20 min. MS/MS of selected ions demonstrated abundant fragment ions and provided a second-dimension "fingerprint" of the complex cellular fraction. Preliminary application of this technique for direct characterization of microorganism lysates is presented.