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Protein A resin for complete elution of Fc binding proteins at a higher pH than existing Protein A resins.
For large-scale purification of antibodies and Fc fusion proteins, Protein A affinity chromatography is the most used capture step in almost every established purification platform. Combining high purity and yield with concentration and rapid transfer into a stable intermediate makes Protein A affinity chromatography the choice. Today, all Protein A resins on the market requires acidic condition (pH 3-4) to achieve a quantitative elution into a reasonable pool volume (2-3 column volumes). This is rarely a problem with most antibodies. However, with the advancement of antibody engineering and drug development, new antibody types such as Fc-fusion proteins and other antibody derived drugs are emerging. This type of molecule is commonly more sensitive to acidic conditions and will in some cases aggregate when typical Protein A elution buffers are used (1). Several publications (2–4) address this by engineering Protein A binding domains for milder elution conditions.
However, to the best of our knowledge, there are currently no commercially available resins that fulfill the performance requirements of a modern Protein A resin while allowing elution at a significantly increased pH. To address this issue, Repligen and Purolite have developed a new type of Protein A resin where complete elution of Fc binding proteins can be achieved at a significantly higher pH compared to existing Protein A resins on the market. In addition to milder elution, a new resin also needs to fulfill other requirements of a process resin described below.
Background: Protein A Affinity Chromatography for Large-Scale Manufacturing
Protein A affinity chromatography has gone through several development phases since the introduction of the first resin intended for large-scale chromatography by Pharmacia (now Cytiva) in 1988. The Protein A itself has been engineered for increased alkaline stability and high capacity and are manufactured without any contact with materials of mammalian origin. The base matrices have been further developed to handle requirements for large-scale batch manufacturing as well as different formats of continuous manufacturing. When looking at the future of Protein A chromatography, one could start from the following checklist of resin properties and their potential impact on downstream processes.......
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Reducing the Costs of Biopharmaceutical Separations and Purifications with Novel Resins in partnership with BioPharm International. Alongside Purolite, YMC America, Process Technologies Group have contributed in sharing their expertise, research and insights in relation to biopharmaceutical separations & purification with novel resins.