Nejvíce citovaný článek - PubMed ID 16996531
Sample preparation involving the cleavage of proteins into peptides is the first critical step for successful bottom-up proteomics and protein analyses. Time- and labor-intensiveness are among the bottlenecks of the commonly used methods for protein sample preparation. Here, we report a fast online method for postinjection acid cleavage of proteins directly in the mobile phase typically used for LC-MS analyses in proteomics. The chemical cleavage is achieved in 0.1% formic acid within 35 s in a capillary heated to 195 °C installed upstream of the analytical column, enabling the generated peptides to be separated. The peptides generated by the optimized method covered the entire sequence except for one amino acid of trastuzumab used for the method development. The qualitative results are extraordinarily stable, even over a long period of time. Moreover, the method is also suitable for accurate and repeatable quantification. The procedure requires only one manual step, significantly decreasing sample transfer losses. To demonstrate its practical utility, we tested the method for the fast detection of ricin. Ricin can be unambiguously identified from an injection of 10 ng, and the results can be obtained within 7-8 min after receiving a suspicious sample. Because no sophisticated accessories and no additional reagents are needed, the method can be seamlessly transferred to any laboratory for high-throughput proteomic workflows.
Biosensors are analytical devices combining a physical sensor with a part of biological origin providing sensitivity and selectivity toward analyte. Biological warfare agents are infectious microorganisms or toxins with the capability to harm or kill humans. They can be produced and spread by a military or misused by a terrorist group. For example, Bacillus anthracis, Francisella tularensis, Brucella sp., Yersinia pestis, staphylococcal enterotoxin B, botulinum toxin and orthopoxviruses are typical biological warfare agents. Biosensors for biological warfare agents serve as simple but reliable analytical tools for the both field and laboratory assay. There are examples of commercially available biosensors, but research and development of new types continue and their application in praxis can be expected in the future. This review summarizes the facts and role of biosensors in the biological warfare agents' assay, and shows current commercially available devices and trends in research of the news. Survey of actual literature is provided.
- Klíčová slova
- Bacillus anthracis, anthrax, bioassay, biological warfare agent, biological weapon, biosensor, colorimetry, electrochemistry, hand held assay, hemorrhagic fever, tularemia,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
