Premium
A simple and inexpensive approach to interfacing high‐performance liquid chromatography and matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry
Author(s) -
Fung Kim Y. C.,
Askovic Srdjan,
Basile Franco,
Duncan Mark W.
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300843
Subject(s) - chromatography , chemistry , mass spectrometry , protein mass spectrometry , capillary electrophoresis–mass spectrometry , matrix (chemical analysis) , matrix assisted laser desorption/ionization , sample preparation , sample preparation in mass spectrometry , desorption , analytical chemistry (journal) , electrospray ionization , adsorption , organic chemistry
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.