Stimuli-Responsive magnetic nanoparticles for monoclonal antibody purification

Luís Borlido, Leila Moura, Ana M. Azevedo, Ana C.A. Roque, Maria R. Aires-Barros*, José P. S. Farinha

*Corresponding author for this work

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Monoclonal antibodies (mAbs) are important therapeutic proteins. One of the challenges facing large-scale production of monoclonal antibodies is the capacity bottleneck in downstream processing, which can be circumvented by using magnetic stimuli-responsive polymer nanoparticles. In this work, stimuli-responsive magnetic particles composed of a magnetic poly(methyl methacrylate) core with a poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) shell cross-linked with N, N'-methylenebisacrylamide were prepared by miniemulsion polymerization. The particles were shown to have an average hydrodynamic diameter of 317 nm at 18°C, which decreased to 277 nm at 41°C due to the collapse of the thermo-responsive shell. The particles were superparamagnetic in behavior and exhibited a saturation magnetization of 12.6 emu/g. Subsequently, we evaluated the potential of these negatively charged stimuli-responsive magnetic particles in the purification of a monoclonal antibody from a diafiltered CHO cell culture supernatant by cation exchange. The adsorption of antibodies onto P(NIPAM-co-AA)-coated nanoparticles was highly selective and allowed for the recovery of approximately 94% of the mAb. Different elution strategies were employed providing highly pure mAb fractions with host cell protein (HCP) removal greater than 98%. By exploring the stimuli-responsive properties of the particles, shorter magnetic separation times were possible without significant differences in product yield and purity.

Original languageEnglish
Pages (from-to)709-717
Number of pages9
JournalBiotechnology Journal
Volume8
Issue number6
Early online date19 Feb 2013
DOIs
Publication statusPublished - Jun 2013
Externally publishedYes

Keywords

  • Cation exchange purification
  • Downstream processing
  • Magnetic particles
  • Monoclonal antibody
  • Stimuli-responsive

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

Fingerprint Dive into the research topics of 'Stimuli-Responsive magnetic nanoparticles for monoclonal antibody purification'. Together they form a unique fingerprint.

  • Cite this

    Borlido, L., Moura, L., Azevedo, A. M., Roque, A. C. A., Aires-Barros, M. R., & Farinha, J. P. S. (2013). Stimuli-Responsive magnetic nanoparticles for monoclonal antibody purification. Biotechnology Journal, 8(6), 709-717. https://doi.org/10.1002/biot.201200329