This Application Note describes a method for studying noncovalent protein interactions using the HPLC-Chip/MS system coupled with an ESI-TOF mass spectrometer. The application note studies apomyoglobin and myoglobin protein complexes. The results show that ESI/MS is an efficient and accurate method for studying noncovalent protein interactions.
This document details a study analyzing noncovalently associated protein complexes, specifically apomyoglobin and myoglobin, using electrospray ionization mass spectrometry (ESI/MS). The study highlights the sensitivity, speed, and selectivity of the ESI/MS approach compared to traditional methods like ultracentrifugation and calorimetry. It demonstrates how the technique can identify protein complexes with high accuracy using small sample amounts.
| Mass accuracy | < 2 ppm |
|---|---|
| Resolution | >12,000 |
| ESI voltage | 1650 V |
| Drying gas flow | 4.0 L/min |
| Drying gas temperature | 350°C |
| Fragmentor voltage | 275-300 V |
| Mass range | 400-3200 Daltons |
| Scan speed | 1 scan/second |
| Infusion flow rate | 600 nL/min |
An Agilent 6210 Time-of-Flight mass spectrometer equipped with an HPLC-Chip MS interface and Infusion Chip was used.
The TOF mass analyzer has a mass accuracy of less than 2 ppm.
The analysis is completed in less than five minutes.
Each sample was infused into the TOF at 600 nL/min using a syringe pump.