RAMOT at Tel Aviv University Ltd.

The Ronit Library-A human synthetic combinatorial antibody library

Posted by RAMOT at Tel Aviv University Ltd.Responsive · Innovative Products and Technologies · Israel

Summary of the technology

The “Ronit 1 Library” is a human synthetic single-chain Fv antibody library of 2?109 independent clones where human complementarity-determining region (CDR) gene fragments of any germ line have been incorporated combinatorially into the appropriate positions of variable-region general frameworks. VH-DP47, VL-DPL3 and VL-DPK22 were used as master frameworks for variable domains of the heavy, lambda light and kappa light chains, respectively. The antibody genes in the library were built on single master frameworks into which diverse CDRs were allowed to recombine. These CDRs were sampled from in vivo-processed gene sequences, thus potentially optimizing the levels of correctly folded and functional molecules, and resulting in a molecule exhibiting a lower immunogenicity compared to naive immunoglobulins.

The repertoire of scFvs represented in the “Ronit 1 library” is cloned into pCC16 phagemid vector. As such, the scFvs may be displayed on phage by infection of the host E. coli with a helper phage or may be expressed as soluble scFvs.

Project ID : 2-2012-348

Description of the technology

The Technology

The “Ronit 1 Library” is a human synthetic single-chain Fv antibody library of 2´109 independent clones where human complementarity-determining region (CDR) gene fragments of any germ line have been incorporated combinatorially into the appropriate positions of variable-region general frameworks. VH-DP47, VL-DPL3 and VL-DPK22 were used as master frameworks for variable domains of the heavy, lambda light and kappa light chains, respectively. The antibody genes in the library were built on single master frameworks into which diverse CDRs were allowed to recombine. These CDRs were sampled from in vivo-processed gene sequences, thus potentially optimizing the levels of correctly folded and functional molecules, and resulting in a molecule exhibiting a lower immunogenicity compared to naive immunoglobulins. 

The repertoire of scFvs represented in the “Ronit 1 library” is cloned into pCC16 phagemid vector. As such, the scFvs may be displayed on phage by infection of the host E. coli with a helper phage or may be expressed as soluble scFvs. 

Potential Applications

 

A robust tool for in-house antibody discovery

Advantages

Human scFv antibody phage display library·         scFvs are fused at their C-termini to the cellulose binding domain (CBD) and can be immobilized onto cellulose either when displayed on phage or when expressed as soluble scFv-CBD fusions

Stage of Development

The “Ronit 1 Library” has been screened against individual peptides and proteins or against protein-peptide complexes and the best selected scFvs exhibited dissociation constants in the subnanomolar range in BIACore analyses.

Reference

JMB (2004) 335, 117-192, "A Human Synthetic Combinatorial Library of Arrayable Single-chain Antibodies based on Shuffling in Vivo Formed CDRs into General Framework Regions

Project manager

Adi Elkeles
BD Manager

Project researchers

Itai Benhar
T.A.U Tel Aviv University, Life Sciences
Molecular Microbiology-Biotechnology

Related keywords

  • Medicine, Human Health
  • Diagnostics, Diagnosis
  • Medical Technology / Biomedical Engineering
  • Virus, Virology / Antibiotics / Bacteriology
  • Biology / Biotechnology
  • Cellular and Molecular Biology Technology
  • Microbiology Technology
  • Bioinformatics Technology
  • Micro- and Nanotechnology related to Biological sciences
  • Monoclonal Antibodies and Hybridomas
  • Microbiology Market
  • Micro- and Nanotechnology related to Biological sciences
  • Biochemistry / Biophysics Market
  • Stem cells and biobanks
  • Cellular and Molecular Biology Market
  • Bioinformatics Market
  • Diagnostic
  • Medical imaging
  • In-vitro diagnostics
  • Therapeutic
  • Other medical/health related (not elsewhere classified)
  • Clinical Medicine
  • Medical equipment
  • oncology
  • Antibodies
  • Life Sciences and Biotechnology
  • Oncology / Cancer
  • In Vitro Diagnostics
  • Pharmaceuticals Indications
  • Medical Devices and Diagnostics
  • Infection Diseases

About RAMOT at Tel Aviv University Ltd.

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