This article discusses the application of mass spectrometry technology in protein research, focusing on the advantages and challenges of electrospray ionization combined with time-of-flight mass spectrometer for protein identification.
This document is an application note by Agilent Technologies exploring the use of electrospray ionization (ESI) coupled with orthogonal-acceleration reflectron time-of-flight mass spectrometry (oa-TOF MS) for identifying intact proteins. It highlights the system's ability to detect small variations in protein structure and identify low-abundance proteins in complex mixtures.
| Column | Poroshell 300SB-C18, 1x75 mm, 5 pm |
|---|---|
| Flow rate (standard) | 0.6 mL/min |
| Injection volume | 1-3 pL |
| Ionization mode | Positive ESI |
| Veap voltage | 4000 V |
| Scan range | m/z 300-2000 |
Experiments were performed using an electrospray ionization (ESI) interface on an Agilent LC/MSD TOF system coupled to an Agilent 1100 Series LC system.
The LC/MSD TOF is capable of rapid scanning, making it compatible with faster chromatographic methods even when proteins are not completely resolved.
Sensitivity decreases by only 20-30% when 90% of the sample is split away prior to analysis.