Over 3000 E-Learning topics / 5000 Articles & Applications
 

Peptide Mapping in a Routine Environment

Once the protein has been fully characterized using LC-MS, an identical method can often be applied for routine peptide mapping in a Good Manufacturing Practice (GMP) production or quality control (QC) environment.

The only substantial difference in the methodology is the use of a variable wavelength UV detector (VWD), typically at 214 nm, rather than a mass spectrometer.

The chromatography is identical to that generated when hyphenated with an MS detector and the UV VWD detector provides more than adequate levels of sensitivity for production and QC purposes.

This is demonstrated in Figure 1 where an isoaspartate peak, resulting from asparagine deamidation of peptide T11 of trastuzumab, is observed at < 0.35% with a signal-to-noise value ‚Č• 35 : 1.

Sensitivity of a trastuzumab peptide map using UV VWD @ 214nm

Figure 1: Sensitivity of a trastuzumab peptide map using UV VWD @ 214nm. 150 x 2.1 mm, 1.8 µm HSS T3, mobile phase A: 0.1% TFA, mobile phase B: 0.1% TFA in acetonitrile, flow rate 300 µL/min, 30 °C, gradient 1-40 %B 0-36.3 minutes

 
loading data
loading data
loading data
loading data
loading data
loading data
loading data
 
 
 
 
 Home | About UsContact Us | SubscribeTerms and Conditions | Advertise | Privacy Policy 

loading data

loading data

loading data

 

loading data


loading data