Structural and performance characteristics of representative anion exchange resins used for weak partitioning chromatography
Autor: | Ranga Godavarti, William F. Daniels, Shaojie Zhang, Jeffrey R. Salm, Tim Iskra, Christopher Gallo, Giorgio Carta |
---|---|
Rok vydání: | 2016 |
Předmět: |
0106 biological sciences
Chromatography Ion exchange Resolution (mass spectrometry) Dimer 010401 analytical chemistry Size-exclusion chromatography Antibodies Monoclonal Reproducibility of Results Chromatography Ion Exchange 01 natural sciences Sensitivity and Specificity 0104 chemical sciences Sepharose Diffusion chemistry.chemical_compound Monomer Adsorption chemistry 010608 biotechnology Mass transfer Materials Testing Anion Exchange Resins Biotechnology |
Zdroj: | Biotechnology progress. 33(2) |
ISSN: | 1520-6033 |
Popis: | Weak partitioning chromatography (WPC) has been proposed for the purification of monoclonal antibodies using an anion exchange (AEX) resin to simultaneously remove both acidic and basic protein impurities. Despite potential advantages, the relationship between resin structure and WPC performance has not been evaluated systematically. In this work, we determine the structure of representative AEX resins (Fractogel® EMD TMAE HiCap, Q Sepharose FF, and POROS 50 HQ) using transmission electron microscopy and inverse size exclusion chromatography and characterize protein interactions while operating these resins under WPC conditions using two mAb monomers, a mAb dimer, mAb multimers, and BSA as model products and impurities. We determine the isocratic elution behavior of the weakly bound monomer and dimer species and the adsorptive and mass transfer properties of the strongly bound multimers and BSA by confocal laser scanning microscopy. The results show that for each resin, using the product Kp value as guidance, salt, and pH conditions can be found where mAb multimers and BSA are simultaneously removed. Isocratic elution and adsorption mechanisms are, however, different for each resin and for the different components. Under WPC conditions, the Fractogel resin exhibited very slow diffusion of both mAb monomer and dimer species but fast adsorption for both mAb multimers and BSA with high capacity for BSA, while the Sepharose resin, because of its small pore size, was unable to effectively remove mAb multimers. The POROS resin was instead able to bind both multimers and BSA effectively, while exhibiting a greater resolution of mAb monomer and dimer species. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:425-434, 2017. |
Databáze: | OpenAIRE |
Externí odkaz: |