Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells

Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells Publisher Pubmed

Eyvazi S^{1, 2} ; Kazemi B³ ; Bandehpour M⁴ ; Dastmalchi S^{2, 5}

Show Affiliations

1. Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2. Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
3. Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
4. Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
5. School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

Source: Immunology Letters Published:2017

Abstract

CD24 is a heavy glycosylated protein that is overexpressed in numerous cancer and cancer stem cells. It is involved in the development, invasion, and metastasis of the cancer cells. CD24 can be considered in targeted cancer therapy as a new target. Here, phage display technology was used for the identification of novel scFv antibodies against CD24. To do so, the CD24 protein was expressed and purified from a stable transgenic CHO cell line. The cells were developed by using a CD24 encoding construct, which targets the 18S rRNA gene. The recombinant CD24 was used in the biopanning process. Four rounds of biopanning were performed with the Tomlinson J library. The polyclonal phage ELISA and DNA sequencing of the selected colonies confirmed that specific binders have been enriched during the biopanning process. Based on the DNA sequencing results, three phage-displaying scFv were obtained and their specificity to CD24 was verified in the monoclonal phage ELISA. The results showed that clone 3 (called Jd3) has the highest affinity to CD24 and also is the most abundant clone (80%) among the three clones. Therefore, Jd3 was selected for further analyses. In order to produce soluble Jd3-scFv, the phage was transfected to E. coli BL21 pLysS. Soluble Jd3 scFv was expressed and purified successfully. The soluble Jd3 scFv showed appropriate affinity to recombinant CD24 in the ELISA analysis. Also, the immunocytochemistry experiment showed that the purified Jd3 scFv can bind to the CD24 expressing A549 cells. So, the scFv may be useful in the targeted delivery of drugs or diagnostic nanoparticles to the CD24 expressing cancer cells. To the best of our knowledge, this study is the first report to identify an scFv antibody against CD24, using the phage display method. © 2017

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Pouyan Aminishakib (1)

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Style	Citing Format
MLA	Eyvazi S, et al.. "Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells." Immunology Letters, vol. 190, no. , 2017, pp. 240-246.
APA	Eyvazi S, Kazemi B, Bandehpour M, Dastmalchi S (2017). Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells. Immunology Letters, 190(), 240-246.
Chicago	Eyvazi S, Kazemi B, Bandehpour M, Dastmalchi S. "Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells." Immunology Letters 190, no. (2017): 240-246.
Harvard	Eyvazi S et al. (2017) 'Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells', Immunology Letters, 190(), pp. 240-246.
Vancouver	Eyvazi S, Kazemi B, Bandehpour M, Dastmalchi S. Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells. Immunology Letters. 2017;190():240-246.
BibTex	@article{ author = {Eyvazi S and Kazemi B and Bandehpour M and Dastmalchi S}, title = {Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells}, journal = {Immunology Letters}, volume = {190}, number = {}, pages = {240-246}, year = {2017} }
RIS	TY - JOUR AU - Eyvazi S AU - Kazemi B AU - Bandehpour M AU - Dastmalchi S TI - Identification of a Novel Single Chain Fragment Variable Antibody Targeting Cd24-Expressing Cancer Cells JO - Immunology Letters VL - 190 IS - SP - 240 EP - 246 PY - 2017 ER -

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