Processing of Microbial Samples Using Protein Ultrafiltration Devices for Identification of Bacteria by Mass Spectrometry

摘要

Greater recovery efficiency for biomolecules using bioreactor devices than that of using conventional biological sample processing. Protocols of the bioreactor sample processing affected the number of unique peptides for the bacterial samples. Protoclol-1 had better recover efficiency than that of Protocol-2 processing. Bacterial Identification and classification at different concentration levels (10~(7)-10~(4) cfu)was correctly assigned for all bacteria and unknown samples processed by the bioreactor devices. Linear relationship was established between number of unique bacterial peptides and TIC-peaks area. Nonlinearity was observed comparing number of unique peptides with that of Sequest generated files. Using commercially available bacterial protein extract was advantageous in determining the efficiency of the bioreactor process and the development of label-free semi-quantitative method for measuring bacterial proteins content in various bacterial samples.