A comprehensive method for modeling and simulating ion exchange chromatography of complex mixtures

Autor(es)

Benedini, Leandro J. ; Furlan, Felipe F. ; Figueiredo, Douglas ; Cabrera-Crespo, Joaquin ; Ribeiro, Marcelo P. A. ; Campani, Gilson ; Gonsalves, Viviane M. ; Zangirolami, Teresa C.

• Tipo de documento: Artigo Científico
• Fonte: Protein Expression and Purification; v. 205, p. 16-pg., 2023-02-13.

Resumo

In recent years, many biological-based products have been developed, representing a significant fraction of income in the pharmaceutical market. Ion exchange chromatography is an important downstream step for the purification of target recombinant proteins present in clarified cell extracts, together with many other unknown impurities. This work develops a robust approach to model and simulate the purification of untagged heterologous proteins, so that the improved conditions to carry out an ion exchange chromatography are identified in a rational basis prior to the real purification run itself. Purification of the pneumococcal surface protein A (PspA4Pro) was used as a case study. This protein is produced by recombinant Escherichia coli and is a candidate for the manufacture of improved pneumococcal vaccines. The developed method combined experimental and computational procedures. Different anion exchange operating conditions were mapped in order to gather a broad range of representative experimental data. The equilibrium dispersive and the steric mass action equations were used to model and simulate the process. A training strategy to fit the model and separately describe the elution profiles of PspA4Pro and other proteins of the cell extract was applied. Based on the simulation results, a reduced ionic strength was applied for PspA4Pro elution, leading to increases of 14.9% and 11.5% for PspA4Pro recovery and purity, respectively, compared to the original elution profile. These results showed the potential of this method, which could be further applied to improve the performance of ion exchange chromatography in the purification of other target proteins under real process conditions. (AU)

• Processo FAPESP: 15/10291-8 - Intensificação e integração de processos de produção e purificação de proteína A de superfície de pneumococo
• Beneficiário: Teresa Cristina Zangirolami
• Modalidade de apoio: Auxílio à Pesquisa - Regular