Peptide Map of biogeneric Alpha Interferon (unreduced) on Kinetex 2.6µm C18 150 x 4.6mm ID

Application Detail (App ID: 18767)

Part No:00F-4462-E0
Dimensions:150 x 4.6 mm ID
Elution Type:Gradient
Elution A:0.1% TFA/2% ACN/Water
Elution B:0.09% TFA/98% ACN/Water
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 99 1
2 40 44 56
3 41 44 56

Flow Rate:1 mL/min
Col. Temp: 25 °C
Detection: UV-Vis Abs.-Variable Wave.(UV) @ 214 nm (22°C)
Note:Application Focus: Peptide mapping with Kinetex C18 for Biogeneric Proteins

Peptide mapping using different proteases is a good analytical method to screen PTMs of therapeutic proteins and is shown on Kinetex 2.6µm C18 for a biogeneric alpha interferon. Interferon-alpha was tryptic digested (1:25 E/S ratio, 18 hrs at 37C in 0.1M NH4HCO3 pH 8.0). Quenched digests (with 1%TFA) were run on the Kinetex 2.6u C18 column (150x4.6) to demonstrate its utility for peptide mapping using a standard method used for a fully porous media.

Results show good resolution for very closely eluting peptides due much in part to the very high efficiency delivered by the Kinetex core shell media. As nearly 15ug of protein digest was loaded on column, the core shell media demonstrate good analytical loading as peak shapes are excellent for nearly every peptide in the map in under 40 minutes despite not being optimized for the media. With proper optimization resolution could be maintained while shortening run time significantly just by increasing flow rate and reducing gradient time to maintain column volume across the gradient.

Analyte Details

Analyte Detail not available

Human alpha Interferon