Rapid optimization of processes for the integrated purification of biopharmaceuticals.

Autor: Crowell LE; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Rodriguez SA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Love KR; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Cramer SM; Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, New York, USA.; Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA., Love JC; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2021 Sep; Vol. 118 (9), pp. 3435-3446. Date of Electronic Publication: 2021 May 04.
DOI: 10.1002/bit.27767
Abstrakt: Straight-through chromatography, wherein the eluate from one column passes directly onto another column without adjustment, is one strategy to integrate and intensify manufacturing processes for biologics. Development and optimization of such straight-through chromatographic processes is a challenge, however. Conventional high-throughput screening methods optimize each chromatographic step independently, with limited consideration for the connectivity of steps. Here, we demonstrate a method for the development and optimization of fully integrated, multi-column processes for straight-through purification. Selection of resins was performed using an in silico tool for the prediction of processes for straight-through purification based on a one-time characterization of host-cell proteins combined with the chromatographic behavior of the product. A two-step optimization was then conducted to determine the buffer conditions that maximized yield while minimizing process- and product-related impurities. This optimization of buffer conditions included a series of range-finding experiments on each individual column, similar to conventional screening, followed by the development of a statistical model for the fully integrated, multi-column process using design of experiments. We used this methodology to develop and optimize integrated purification processes for a single-domain antibody and a cytokine, obtaining yields of 88% and 86%, respectively, with process- and product-related variants reduced to phase-appropriate levels for nonclinical material.
(© 2021 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals LLC.)
Databáze: MEDLINE
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