Abstract
The analysis of monoclonal antibodies (mAbs) by a middle-down approach is a growing field that attracts the attention of many researchers and biopharma companies. Usually, liquid fractionation techniques are used to separate mAbs polypeptides chains before mass spectrometry (MS) analysis. Gas-phase fractionation techniques such as high-field asymmetric waveform ion mobility spectrometry (FAIMS) can replace liquid-based separations and reduce both analysis time and cost. Here, we present a rapid FAIMS tandem MS method capable of characterizing the polypeptide sequence of mAbs light (Lc) and heavy (Hc) chains in an unprecedented, easy, and fast fashion. This new method uses commercially available instruments and takes ∼ 24 minutes —40-60% faster than regular LC-MS/MS analysis — to acquire fragmentation data using different dissociation methods.
Abbreviations
- mAb
- Monoclonal Antibody
- IgGs
- Immunoglobulins G
- Lc
- Light Chain
- Hc
- Heavy Chain
- FDA
- Food and Drug Administration
- MS
- Mass Spectrometry
- LC–MS/MS
- Liquid Chromatography Tandem Mass Spectrometry
- S/N
- Signal to Noise
- MS/MS
- Tandem Mass Spectrometry
- RP
- Reverse Phase
- SEC
- Size Exclusion
- IEX
- Ion Exchange
- FAIMS
- High-field asymmetric waveform ion mobility spectrometry
- DC
- Direct Current
- CV
- Compensation Voltage
- TECP-HCl
- (2-carboxyethyl)phosphine Hydrochloride
- AGC
- Automatic Gain Control
- CID
- Collision Induced Dissociation
- DV
- Dispersion Voltage
- HCD
- Higher-energy Collisional Dissociation
- NCE
- Normalized Collision Energy
- UVPD
- Ultraviolet Photodissociation
- ETD
- Electron Transfer Dissociation
- EThc
- DElectron-transfer/Higher-energy Collision Dissociation
- Hex
- Hexose
- PTMs
- Post-translational Modifications