Hajarsadat Ghaderi - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Zahra Noormohammadi - Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Mahdi Habibi-Anbouhi - National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran
Fatemeh Kazemi-Lomedasht - Biotechnology Research Centre, Venom and Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, Iran
Mahdi Behdani - Biotechnology Research Centre, Venom and Biotherapeutics Molecules Laboratory, Pasteur Institute of Iran, Tehran, Iran

Objective(s): SLC۳۹A۶ (solute carrier family ۳۹) or LIV-۱, is a zinc-transporter protein associated with estrogen-positive breast cancer and its metastatic spread. Significantly there is a direct relation between high zinc intake and unregulated proliferation and cancers. Blocking SLC۳۹A۶ protein may result in reduced metastasis and proliferation in many malignant tumors. This study aimed to develop an anti-SLC۳۹A۶ nanobody that is able to detect and block the SLC۳۹A۶ protein on the surface of cancerous cells.Materials and Methods: The recombinant SLC۳۹A۶ was expressed and used for camel immunization. The VHH library was constructed and screened for SLC۳۹A۶-specific nanobody. Then, the strength of nanobody in SLC۳۹A۶ detection was evaluated by Western blotting and flow cytometry.Results: We showed the ability of nanobody to detect SLC۳۹A۶ by Western blotting and flow cytometry and the specificity of the C۳ nanobody for the SLC۳۹A۶ antigen. Furthermore, the selected nanobody potently inhibits cell proliferation.Conclusion: These data show the potential of SLC۳۹A۶-specific nanobody for the blockade of zinc transportation and provide a basis for the development of novel cancer therapeutics.

Breast Cancer, Nanobody, Phage display, SLC۳۹A۶, VHH library