Abstract:
:The ability to obtain the accurate mass of a protein in a complex sample mixture aids in determining its correct in vivo form. This is important when identifying post-translationally modified proteins, protein variants or isoforms. The central technique used to separate proteins, 2-dimensional gel electrophoresis offers excellent separation capabilities but does not provide adequate mass accuracy. In this study, an alternative method, liquid chromatography (LC) coupled with matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF)-MS (LC-MALDI) is described. LC-MALDI-MS was used to separate and determine the mass of proteins and peptides in a complex biological sample (i.e., human pituitary gland homogenate). Peptides and proteins were first separated by capillary chromatography and the eluent mixed post-column with sinapinic acid matrix. The flow was then deposited directly onto a standard MALDI target via a capillary nebulizer. In addition to offering high mass accuracy, this method can be applied to peptide and protein quantification.
journal_name
Proteomicsjournal_title
Proteomicsauthors
Fung KY,Askovic S,Basile F,Duncan MWdoi
10.1002/pmic.200300843subject
Has Abstractpub_date
2004-10-01 00:00:00pages
3121-7issue
10eissn
1615-9853issn
1615-9861journal_volume
4pub_type
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