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pH Elution Properties of Purolite™ Affinity Resins/Blog/Ecolab
Application Note
pH Elution Properties of Purolite™ Affinity Resins
Overview
Protein A affinity resins play a crucial role in purifying monoclonal antibodies (mAbs) and related therapeutic molecules, serving as the most reliable and efficient initial step in downstream purification.
During the purification step, after the target molecule binds to the affinity column, the pH is lowered to achieve elution. However, some mAbs are damaged at low pH levels, while other antibodies, including bispecifics, show high aggregation. For such molecules, a mild pH elution is particularly beneficial.
This study compares the pH elution of two market-leading resins with two Purolite protein A resins, one specifically designed for high pH elution.
Figure 1 pH Elution Profile ComparisonsComparison of four resins using a standard monoclonal antibody: Purolite Praesto Jetted A50 (blue), Purolite Praesto Jetted A50 HipH (teal), Competitor A (red), and Competitor B (green).
Investigation
In this study, the pH elution profiles of two Purolite resins were investigated and compared to two market-leading protein A resins.
The first Purolite chromatography resin, Purolite™ Praesto™ Jetted A50, is a native protein A resin, designed to purify a wide range of mAb constructs. The second resin, Purolite™ Praesto™ Jetted A50 HipH, allows for the elution of the bound monoclonal antibody molecule at a higher pH. This resin is particularly suited for complex purification procedures, including pH-sensitive antibodies prone to aggregation at typical pH elution levels.
To form a comparison, two competitor resins were also investigated. Competitor A is a widely used native protein A resin, and Competitor B is an engineered high-performance protein A resin.
Conclusion
The study showed that three of the resins (Purolite Praesto Jetted A50, Competitor A, Competitor B) eluted around pH 4 or below. However, Purolite Praesto Jetted HipH demonstrated a considerably higher pH elution.
The high pH elution of Purolite Praesto Jetted A50 HipH is a key feature of the resin, designed into the ligand to allow for maximum recovery of the target molecule when aggregation proves challenging, particularly with bispecific antibodies and other complex structures with multiple functionalities.
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