Magnetic Nanoparticle-Based Semi-automated Panning for High-Throughput Antibody Selection

Methods Mol Biol. 2023:2702:291-313. doi: 10.1007/978-1-0716-3381-6_15.

Abstract

Bio-panning is a common process involved in recombinant antibody selection against defined targets. The biopanning process aims to isolate specific antibodies against an antigen via affinity selection from a phage display library. In general, antigens are immobilized on solid surfaces such as polystyrene plastic, magnetic beads, and nitrocellulose. For high-throughput selection, semi-automated panning selection allows simultaneous panning against multiple target antigens adapting automated particle processing systems such as the KingFisher Flex. The system setup allows for minimal human intervention for pre- and post-panning steps such as antigen immobilization, phage rescue, and amplification. In addition, the platform is also adaptable to perform polyclonal and monoclonal ELISA for the evaluation process. This chapter will detail the protocols involved from the selection stage until the monoclonal ELISA evaluation with important notes attached at the end of this chapter for optimization and troubleshooting purposes.

Keywords: Antibody library; Magnetic nanoparticle; Monoclonal antibodies; Panning; Phage display; Semi-automated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies
  • Bacteriophages*
  • Bioprospecting
  • Cell Surface Display Techniques
  • Humans
  • Magnetite Nanoparticles*

Substances

  • Magnetite Nanoparticles
  • Antibodies