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

应用详细信息 (应用ID: 18767)

尺寸:150 x 4.6 mm ID
洗脱A:0.1% TFA/2% ACN/Water
洗脱B:0.09% TFA/98% ACN/Water
步骤序号 时间 (分) 图片A 图片B
1 0 99 1
2 40 44 56
3 41 44 56

流速 (毫升/分钟):1 mL/min
冷却温度: 25 °C
检测: UV-Vis Abs.-Variable Wave.(UV) @ 214 nm (22°C)
备注: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.



Human alpha Interferon