( ...)Using the Agilent 2100 bioanalyzer for,,,quality control of protein samples prior to,,,MS-analysis

Whereas in the past mass spectrometers (MS) were confined to the realm of small molecules, the development of soft ionization methods such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) revolutionized the analysis of large biomolecules1, 2. Today, the analysis of proteins up to 300 kDa using MS has become a routine method to gain molecular weight information with high accuracy and structural information. However, sample purity greatly influences the results of MS analysis. In MS service facilities, scientists have to rely on the information provided by the customers and the quality of the sample preparation they have done. Bad results are often the consequence of customers being too optimistic about their own samples. Especially concentration and purity are often overestimated. To avoid unproductive MS runs, a fast and simple precheck is valuable. In this Application Note we show that the Agilent 2100 bioanalyzer can be used for quality control of protein samples prior to MS analysis. A corresponding data set analyzed with LC/MS and with the Agilent 2100 bioanalyzer using the Protein 200 Plus LabChip kit is shown.
Experiments
All samples were adjusted to a concentration of 1 mg/mL by dilution with the appropriate buffer prior to the analysis (based on information given by the customer).
LC-MS analysis
The experimental setup comprised:
Micro-HPLC-Pump (ABI 140C)
Autosampler (PE200)
HPLC column (Nucleosil C8 125 x 2 mm, 5 , 120 from Macherey & Nagel)
UV detector (ABI 785A) set to 214 nm, in row with
Single quadrupole mass spectrometer (API165 from Applied Biosystems) wi
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