A Comparison Between Cell, Protein and Peptide-Based Approaches for Selection of Nanobodies Against CD44 from a Synthetic Library

Title:A Comparison Between Cell, Protein and Peptide-Based Approaches for Selection of Nanobodies Against CD44 from a Synthetic Library

Volume: 25 Issue: 6

Author(s): Soudabeh Kavousipour, Pooneh Mokarram*, Seyed Latif Mousavi Gargari, Zohreh Mostafavi-Pour, Mehdi Barazesh, Amin Ramezani, Hassan Ashktorab, Shiva Mohammadi and Saeid Ghavami

Affiliation:

• Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz,Iran

Keywords: Cell-based panning, CD44, Nanobody, Phage display, VHH, Synthetic library.

Abstract: Background: The hyaluronic acid receptor CD44, is a cancer stem cell biomarker, playing important roles in cell adhesion, tumor progression and drug-resistance. Therefore, CD44 is a potential target for cancer treatment and its blockade could result in multi-factorial therapeutic effects.

Methods: Nanobodies against CD44 were isolated from a synthetic library with a diversity of 5×1011 CFU/ml using the phage display technique. Three approaches were used for isolation of nanobodies fragments including peptide-, protein- and cell-based panning.

Results: Nanobodies from cell-based panning displayed more specificity compared to protein or peptide-based panning. Our results show that cell-based panning is the most efficient method for isolation of a specific single domain antibody fragment to CD44 from a synthetic phage displayed library.

Conclusion: The isolated nanobodies could successfully recognize and bind cells that express the CD44 surface antigen.

Cite this article as:

Kavousipour Soudabeh , Mokarram Pooneh *, Gargari Latif Mousavi Seyed , Mostafavi-Pour Zohreh , Barazesh Mehdi , Ramezani Amin , Ashktorab Hassan , Mohammadi Shiva and Ghavami Saeid , A Comparison Between Cell, Protein and Peptide-Based Approaches for Selection of Nanobodies Against CD44 from a Synthetic Library, Protein & Peptide Letters 2018; 25 (6) . https://dx.doi.org/10.2174/0929866525666180530122159