Monitoring virus-like particle and viral protein production by intact cell MALDI-TOF mass spectrometry
| Autor: | Paula M. Alves, Maria Candida M. Mellado, Catarina Franco, Ana Varela Coelho |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Cell Nucleus
Chromatography medicine.diagnostic_test Viral protein Chemistry Cell medicine.disease_cause Mass spectrometry Mass Spectrometry Analytical Chemistry law.invention Viral Proteins Matrix-assisted laser desorption/ionization medicine.anatomical_structure Virus-like particle Western blot law Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization medicine Recombinant DNA Intracellular |
| Zdroj: | Talanta. 80:1561-1568 |
| ISSN: | 0039-9140 |
| DOI: | 10.1016/j.talanta.2009.06.081 |
| Popis: | A new application of intact cell MALDI-TOF MS (ICM-MS) methodology is described for monitoring the production of viral proteins and viral like particles using the baculovirus/insect cells expression system. Various MALDI matrices, cell preparation methods, cell/matrix volume ratio and MALDI target application procedures were tested in order to obtain the highest intensity and reproducibility of intact insect cell spectra. The web interface, SPECLUST (http://bioinfo.thep.lu.se/speclust.html), was used to construct dendograms based on MALDI-TOF MS data for evaluation of fingerprint changes. We demonstrate that insect cell mass spectrum fingerprints are characteristic of each viral protein/particle production. Their changes along the time for each production experiment correlate with the intracellular viral protein content determined by Western blot. This work shows that this simple, fast and low cost assay, which requires low sample volume, is a powerful analytical tool that complements the most common analytical methods used for monitoring bioprocesses and has potential application in the biotechnological industry namely, in the production of recombinant proteins. |
| Databáze: | OpenAIRE |
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