Application of phage display (Part 1)-Special Topic on Phage Display Technology-Tekbiotech-Yeast and Phage Display CRO, Expert in Nano-body and Antibody Drug Development

Application of phage display (Part 1)

Application 1: Therapeutic Antibody Development

1.1 Antibody Discovery Against Difficult Targets

The conventional therapeutic development workflow starts with target identification, followed by antibody generation against the target. However, not all targets are readily tractable. Many targets are unsuitable for traditional hybridoma platforms due to high sequence conservation, intrinsic toxicity, membrane protein properties, multiple transmembrane domains, poor immunogenicity, or absence of characterized functional epitopes.

Powered by PhagePro Tech^?, in vitro biopanning bypasses animal immunization via de novo synthetic antibody libraries independent of in vivo immune responses. This platform enables antibody screening against toxic, highly conserved and poorly immunogenic targets. For hard-to-purify antigens, phage display libraries support whole-cell panning and DNA-based panning strategies. Moreover, phage library screening supports directed evolution: site-directed mutagenesis or error-prone mutagenesis can be applied to optimize affinity and thermal stability of lead antibodies derived from any upstream workflow.

1.2 Antibody Humanization

Traditional antibody humanization grafts murine complementarity-determining regions (CDRs) onto human antibody frameworks, which frequently results in compromised antigen binding affinity. PhagePro Tech^? delivers two optimized humanization pipelines to overcome this limitation:

Pipeline 1: Generate murine monoclonal antibodies via hybridoma first; graft CDR3 onto human antibody frameworks. Random mutagenesis is then performed on CDR3 to construct phage libraries. Multiple rounds of antigen-based biopanning and directed evolution yield affinity-matured humanized antibodies.

Pipeline 2: Nearly eliminates full-length CDR grafting. After obtaining murine mAbs, key antigen-binding motifs are identified and embedded into human antibody libraries for antigen-directed screening. The resulting antibodies retain murine antibody specificity while maintaining robust affinity in human contexts, as the majority of framework sequences are human-derived.

Workflow of Antibody Humanization-1.png

Figure 1 Workflow of Antibody Humanization【1】

Application 2: Research on Protein–Protein Interactions

2.1 Mimotope Identification (Core for Vaccine Development)

Mimotope screening is utilized when neutralizing mAbs are obtained from hybridoma platforms, yet the exact virus-antibody binding epitope remains uncharacterized—critical information for broad-spectrum vaccine design. In brief, purified neutralizing antibodies are used to screen phage random peptide libraries. Phages displaying peptide sequences highly homologous to viral epitopes are defined as mimotopes, which serve as templates for epitope-focused vaccine design.

This workflow forms a closed-loop R&D pipeline distinct from de novo target antibody discovery:

Identification of core neutralizing epitopes → Human antibody library panning against epitopes → Generation of therapeutic antibodies

Neutralizing antibody generation via PhagePro Tech? or alternative platforms → Mimotope mapping → Broad-spectrum vaccine design

Mimotope Identification and Translational Applications-2.png

Figure 2 Mimotope Identification and Translational Applications【2】

2.2 Partner Protein Screening

No protein functions independently in biological pathways; identification of interacting partner proteins is fundamental to decoding molecular regulatory networks. PhagePro Tech? enables precise capture of protein binding partners through phage display cDNA library construction.

Briefly, the protein of interest (POI) is immobilized and incubated with phage cDNA libraries. Binding phage clones are isolated, sequenced, and analyzed to identify interacting partner proteins. POIs applicable to this platform include enzymes, signaling molecules, intracellular receptor domains, viral proteins, and any protein of research interest. This technology directly retrieves full-length genes of interacting partners with ultra-high sensitivity and supports functional screening.

Based on proprietary phage display technology, Tech Biotech (Tianjin) Co., Ltd. has established a comprehensive target-specific therapeutic antibody discovery platform. We provide high-quality VHH and neutralizing monoclonal antibody development services for researchers worldwide, covering preclinical discovery and validation for GPCR targets, oncogenic targets, and disease-related targets. Our one-stop technical services span project scheme design to developability assessment, fully meeting global demands for therapeutic monoclonal antibody development.

reference documentation：

[1] Chen HC,Pan YL,Chen Y, et al. Monoclonal Antibodies as a Therapeutic Strategy against Multidrug-Resistant Bacterial Infections in a Post-COVID-19 Era. Life (Basel). 2024;14 (2):.

[2] Luzar J,?trukelj B,Lunder M. Phage display peptide libraries in molecular allergology: from epitope mapping to mimotope-based immunotherapy. Allergy. 2016;71 (11):1526-1532.

Previous: Phage Display Next:Application of phage display (Part 2)