Intact mass analysis by HRMS
Mass spectrometry (MS) is an analytical technique that measures the mass-to-charge ratio of charged particles.
- Molecular weight determination
- Structural characterization
- Qualitative and quantitative analysis of components in a mixture.
Areas of applications:
- Pharmaceutical analysis
- Forensic analysis
- Environmental analysis
- Clinical applications
ORBITRAP MASS SPECTROMETRY
Orbitrap mass spectrometers deliver a total possible maximum resolution (FWHM) of 1,000,000 at m/z 200 and a sub-1 ppm mass accuracy in a single compact and easy-to-use instrument. These high-resolution accurate-mass systems detect a wide range of compounds and small molecules during both targeted and untargeted analyses, without losing selectivity or sensitivity. Simply put, when it comes to Orbitrap technology, there is no compromise.
High Resolution Mass Spectrometer (HRMS) AT CBA FACILITY
Make: ThermoFisher Scientific
Model: Q Exactive Plus Biopharma Orbitrap Mass Spectrometer
- Resolution up to 240,000 (FWHM) at m/z 200
- Analysis of large molecules up to m/z 8000
INTACT MASS ANALYSIS OF PROTEINS
- Mass analysis of intact proteins using high resolution mass spectrometry is a rapid method for confirming the identity and the primary structure of proteins.
- Mass analysis can provide information for evaluating and comparing the isoforms, glycoforms, and molecule wide modifications associated with a protein.
- The analysis can readily compare manufacturing batches or can be used to compare a biosimilar to the innovator.
- Measurement of molecule weight is a requirement of the regulatory guidelines.
INTACT MASS –UNDER NON REDUCING CONDITIONS
Figure 2. Raw m/z spectrum of NIST mAb
Figure 3. Deconvoluted Mass (Da) spectra of NIST mAb
The charge envelopes in this spectrum are centered on ions having a charge (z) of 53. Each charge envelope is rich in information about the structure. The NIST mAb has four major glycoforms that can be observed in the inset of Figure 3.
INTACT MASS –UNDER REDUCING CONDITIONS
INTACT MASS ANALYSIS AT CBA
|Intact mass analysis of protein|
|Sample requirement||Amount- 0.1-0.5 ml (1mg/ml)|
|Deliverables||Raw mass spectrum and Deconvoluted mass spectra|
|Information required from Client||Expected molecular weight and formula
Sample solubility, polarity and formulation buffer
Protein sequence in FASTA format for comparison