Arg-SEC Mobile Phase
Effective separation of protein conjugates, e.g., ADC and sticky cytokines / Universal mobile phase for size-exclusion chromatography
Size-Exclusion Chromatography (SEC) is an essential technology for developing biopharmaceuticals. SEC is normally carried out in aqueous solution so that the mobile phase has a minimal impact on the target biopharmaceutical proteins, e.g., not disrupting the protein structure. However, such mobile phase is not often optimal for the performance of SEC. Proteins often bind to the SEC column surface, leading to a compromised protein elution and peak resolution. Such binding can cause incorrect determination of the size of the eluted proteins and the amount of the aggregated species. To overcome these problems, arginine-based mobile phase*1 has been developed. This mobile phase reduces non-specific protein binding to many SEC columns, while not affecting the protein structure. Depending on the strength of non-specific binding, there are 3 mobile phases: Strong, Standard and Mild.
This product is manufactured with permission from Ajinomoto Co., Inc. based on the patent US 7501495*2.
*1 Arginine is effective in suppressing non-specific molecular interactions.
*2 JP: 4941882, US: 7501495, EP: 1698637
Features
- Increased recovery of proteins and peptides that are more hydrophobic and have stronger tendency to stick to the columns.
- Correct determination of protein aggregates that have stronger tendency to stick to the columns than the monomeric species.
- Optimal for quality control.
- Effective separation of protein conjugates, e.g., ADC and sticky cytokines.
Advantages of using arginine-based mobile phase
- Quantitative analysis of protein (in particular sticky protein) samples.
- Applicable to many commercial SEC columns.
- Reduce lot-to-lot variability due to non-specific protein binding.
- Less non-specific protein binding increases the life time of the columns.
- Reduce the time required to condition the columns.
- Increase the purification yield as a result of less binding.
Arg-SEC Mobile Phase Series
| Product ID | Application | Example |
|---|---|---|
| Standard | First choice, all proteins | IgGs |
| Strong | More hydrophobic, sticky proteins | TGF-β3, Aggregates |
| Mild | Oligomeric proteins with weak subunit-subunit interaction | Hemoglobin* |
Filtered through 0.22 µm filter, ready to use, pH 6.8, contains arginine.
*Such a weakly associating oligomeric protein as hemoglobin may dissociate to monomers in the presence of arginine.
Reference:
Mobile Phase Containing Arginine Provides More Reliable SEC Condition for Aggregation Analysis
Journal of Pharmaceutical Sciences 99(2), 618-620 (2010)
Arginine as an effective additive in gel permeation chromatography
Journal of Chromatography A 1094(1-2), 49-55 (2005)
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