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

[ Vol. 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   Pages 580 - 588 ( 9 )

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.

Keywords:

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

Affiliation:

Department of Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Department of Biology, Faculty of Basic Science, Shahed University, Tehran, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Department of Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Department of Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Department of Medicine and Cancer Center, Howard University College of Medicine, Washington, DC, Department of Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, Winnipeg, MB

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