Phage Display‐Derived Antibodies: Application of Recombinant Antibodies for Diagnostics

Antibodies are produced by the human body in response towards infections as a means of protection. The in vivo production of antibodies by B-cells involves a series of intricate gene editing processes resulting in a highly diverse pool of antibodies. However, this diversity can be replicated in vitro using phage display. Phage display offers the potential to present the antibody phenotype together with the cloned genotype of the specific antibody in a single-phage particle. Antibodies are highly sought after for diagnostic applications owing to its specificity and affinity towards a target antigen. The advent of recombinant antibody (rAb) technology allows for a faster and more cost-effective solution for antibody generation. It also provides diagnostic developers with the possibility to customize the antibodies. Antibodies have been utilized successfully in various diagnostic platforms ranging from standard immunoassays to lateral-flow assays, nanoparticles, microfluidics, DNA‐integrated assays and others. The limitless application of antibodies in the field of diagnostics has made it a critical component in any diagnostic development platform. This chapter focuses on the processes involved in antibody discovery including the various forms of antibody libraries for phage display and panning processes. We also highlight some diagnostic platforms that apply recombinant antibodies

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

Abstract Antibody phage display is a key tool for the development of monoclonal antibodies against various targets. However, the development of anti-peptide antibodies is a challenging process due to the small size of peptides for binding. This makes anchoring of peptides a preferred approach for panning experiments. A common approach is by using streptavidin as the anchor protein to present biotinylated peptides for panning. Here, we propose the use of recombinant expression of the target peptide and an immunogenic protein as a fusion for panning. The peptide inhibitor of trans-endothelial migration (PEPITEM) peptide sequence was fused to the Mycobacterium tuberculosis (Mtb) α-crystalline (AC) as an anchor protein. The panning process was carried out by subtractive selection of the antibody library against the AC protein first, followed by binding to the library to PEPITEM fused AC (PEPI-AC). A unique monoclonal scFv antibodies with good specificity were identified. In conclusion, the use of an alternative anchor protein to present the peptide sequence coupled with subtractive panning allows for the identification of unique monoclonal antibodies against a peptide target