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Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
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A Comparison Between Cell, Protein and Peptide-Based Approaches for Selection of Nanobodies Against CD44 from a

Soudabeh Kavousipour1, Pooneh Mokarram2,3, Seyed Latif Mousavi Gargari4

  • 1Department of Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.

Protein and Peptide Letters
|June 1, 2018
PubMed
Abstract

Insights

Researchers developed specific nanobodies targeting CD44, a key marker in cancer stem cells. Cell-based panning proved most effective for isolating these nanobodies, which successfully bind to CD44-expressing cells.

Area of Science:

  • Biotechnology
  • Immunology
  • Oncology

Background:

  • CD44 is a hyaluronic acid receptor and a cancer stem cell biomarker.
  • It plays critical roles in cell adhesion, tumor progression, and drug resistance.
  • Targeting CD44 offers potential for multi-factorial cancer therapeutics.

Purpose of the Study:

  • To isolate and characterize nanobodies against the CD44 receptor.
  • To evaluate different panning strategies for nanobody isolation.

Main Methods:

  • Utilized phage display technique with a synthetic library (5×10^11 CFU/ml).
  • Employed three panning approaches: peptide-based, protein-based, and cell-based.
  • Isolated single-domain antibody fragments (nanobodies) against CD44.

Main Results:

  • Cell-based panning yielded nanobodies with higher specificity compared to peptide or protein-based methods.
  • Demonstrated cell-based panning as the most efficient strategy for isolating specific CD44 nanobodies.
  • Confirmed the ability of isolated nanobodies to recognize and bind CD44-expressing cells.

Conclusions:

  • Successfully isolated nanobodies targeting the CD44 receptor.
  • Cell-based panning is a superior method for generating specific nanobodies against cell surface targets like CD44.
  • The developed nanobodies have potential applications in cancer diagnostics and therapeutics.
Keywords:
CD44Cell-based panningNanobodyPhage displaySynthetic libraryVHH.

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