Affiliations 

  • 1 Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia
  • 2 Max Planck Institute of Colloid and Interfaces, Department of Systems Biology, Potsdam, Germany
  • 3 Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia; Analytical Biochemistry Research Centre, Universiti Sains Malaysia, Penang, Malaysia. Electronic address: theamsoon@usm.my
Methods Enzymol, 2020;630:159-178.
PMID: 31931984 DOI: 10.1016/bs.mie.2019.10.023

Abstract

Directed evolution is a proven approach to fine tune or modify biomolecules for various applications ranging from research to industry. The process of evolution requires methods that are capable of not only generating genetic diversity but also to distinguish the variants of desired characteristics. One method that is synonymous with directed evolution of proteins is phage display. Here, we present a protocol describing the application of magnetic nanoparticles coupled with a processor to carry out the identification of monoclonal antibodies (mAbs) from a diverse antibody library via phage display. Target antigens are coupled to magnetic nanoparticles as the solid phase for the isolation of the binding mAbs via affinity. A gradual enrichment in clones would result in increasing ELISA readouts with increasing rounds of panning. During monoclonal level analysis, positivity can be deduced with comparison to background and controls. The biopanning process can also be adopted for the directed evolution of enzymes, scaffold proteins or even peptides.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.