Recombinant-Chemical Platform for Bispecific scFvs

Overview

This cross-disciplinary, industry-focused project aims to develop a recombinant–chemical platform for producing bispecific single-chain variable fragments (bs-scFvs) using the flashBAC™ baculovirus expression system to produce scFv and conjugation-ligation to prepare bs-scFv. .

The work integrates protein engineering, advanced expression technologies, and site-specific chemical conjugation to accelerate the development of next-generation biotherapeutics. The flashBAC™ system enables rapid, high-yield production of scFvs while maintaining their structural integrity and activity in a cost-effective, scalable manner. By combining recombinant expression with precise chemical assembly, the project establishes a modular workflow to generate bispecific constructs with defined architectures and enhanced therapeutic potential. Collaboration with industry partners ensures the platform is aligned with manufacturing needs and translation to clinical applications.

Through this integrated strategy, the project seeks to demonstrate a streamlined and economically viable workflow for the development of bispecific scFvs, laying the foundation for future innovation in targeted biologics.

Objectives

• Express monospecific scFvs using flashBAC™ baculovirus platform
• Develop site-selective conjugation chemistry for bs-scFv assembly
• Retain antigen binding affinity and structural stability post-ligation
• Enable rapid prototyping of therapeutic formats
• Align the platform with industry-standard workflows for scale-up and translation

Why this matters

Traditional methods of generating bispecific antibodies are labor-intensive, expensive, and poorly scalable. Our integrated approach:

• Reduces development time and cost
• Ensures precise control over bispecific structure and function
• Allows plug-and-play flexibility for targeting multiple antigens
• Facilitates translation from research to manufacturing

Collaboration & Impact

This project is co-developed with industry partners to ensure applicability in biotherapeutic manufacturing. It contributes to THRIVE’s broader mission to deliver efficient, next-generation biologics by bridging academic innovation with commercial readiness.

Technical Framework

• Protein Engineering: Optimised scFv sequence design and construct validation
• Recombinant Expression: High-yield scFv production using flashBAC™
• Chemical Ligation: Controlled, bioorthogonal coupling to form bispecifics
• Characterisation: Affinity, stability, and structural analysis of bs-scFvs

Publications

• J. Toor , WR. Grabowska, A. L. Johnson, J. Jones, WG. Stetler-Stevenson, H. Khalili, D. Peeney. Histidine Tag-Specific PEGylation Improves the Circulating Half-Life of TIMP2. ACS Appl Bio Mater., 2025 Feb 21.

• S. Prikalkhoran, D. Guiliano, H. Khalili, “Storage stability and solution binding affinity of an Fc-fusion mimetic” Journal of Pharmaceutical Sciences, 2025, 114, 2, 1061-1067.

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This project exemplifies translational biotherapeutic research at THRIVE—blending precision chemistry, synthetic biology, and industry collaboration for scalable antibody innovation.