Table 1 Representative methodologies for experimental affinity screening systems

Phage Display

Antibody fragments are displayed on phage coat proteins, enabling selection against immobilized antigens

High throughput, large library sizes, amenable to in vitro evolution

May require specialized equipment, potential to obtain false positives due to phage surface interactions

 < 10¹¹

Yeast Display

Antibodies are displayed on the surface of yeast cells, allowing FACS-based sorting for antigen binding

Eukaryotic protein folding, high throughput screening, amenable to genetic manipulation

Limited by the size of the yeast cell surface, requires specific yeast strains

 < 10⁹

Mammalian Cell Display

Antibodies are expressed on the surface of mammalian cells, providing a native-like environment for screening

Accurate representation of antibody function, allows for post-translational modifications

Lower throughput than phage or yeast display, requires specialized cell lines

 < 10⁸

Ribosome Display

Antibody-mRNA complexes are formed and stabilized on ribosomes, allowing for selection based on antibody-antigen binding

Cell-free system, high throughput, large library sizes, suitable for toxic or difficult-to-express proteins, amenable to in vitro evolution

mRNA-ribosome-protein complexes can be unstable and prone to dissociation during selection. mRNA may be degraded by nucleases in the reaction mixture

 < 10¹⁵