Engineering an anti-granulocyte colony stimulating factor receptor nanobody for improved affinity

(2020) Engineering an anti-granulocyte colony stimulating factor receptor nanobody for improved affinity. Life Sciences. ISSN 0024-3205

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Abstract

Aims: Granulocyte colony-stimulating factor (G-CSF) is a cytokine that induces proliferation and differentiation of hematopoietic precursor cells and activation of mature neutrophils. G-CSF is overexpressed in several malignant tumors and blocking its binding to the receptor can lead to significant decrease in tumor growth, vascularization and metastasis. Furthermore, targeting G-CSF receptor has shown therapeutic benefit in other diseases such as rheumatoid arthritis, progressive neurodegenerative disorder and uveitis. Camelid single-chain antibodies (nanobodies) have exceptional properties making them appropriate for tumor imaging and therapeutic application. In this study we aim to use the rational design approach to engineer a previously described G-CSF-R targeting nanobody (VHH1), to improve its affinity toward G-CSF-R. Main methods: We redesigned the complementary determining region 3 (CDR3) domain of the VHH1 nanobody to mimic G-CSF interaction to its receptor and developed five new engineered nanobodies. Binding affinity of the engineered nanobodies was evaluated by ELISA (Enzyme-linked immunosorbent assay) on NFS60 cells. Key findings: Enzyme-linked immunosorbent assay (ELISA) confirmed the specificity of the engineered nanobodies and ELISA-based determination of affinity revealed that two of the engineered nanobodies (1c and 5a) bind to G-CSF-R on the surface of NFS60 cells in a dose-dependent manner and with a higher potency compared to the parental nanobody. Significance: Additional studies are required to better characterize these nanobodies and assess their interaction with G-CSF-R in vitro and in vivo. These newly developed nanobodies could be beneficial in tumor imaging and therapy and make a basis for development of additional engineered nanobodies.

Item Type: Article
Keywords: Engineered nanobodies G-CSF G-CSF receptor FACTOR G-CSF GM-CSF THERAPEUTIC ANTIBODIES CELL-LINE EXPRESSION IDENTIFICATION ANGIOGENESIS MOBILIZATION CARCINOMA TARGETS
Subjects: QV Pharmacology
QZ Pathology > QZ 200-380 Neoplasms
Divisions: Faculty of Pharmacy and Pharmaceutical Sciences > Department of Pharmaceutical Biotechnology
Isfahan Pharmaceutical Sciences Research center
Journal or Publication Title: Life Sciences
Journal Index: ISI
Volume: 257
Identification Number: https://doi.org/10.1016/j.lfs.2020.118052
ISSN: 0024-3205
Depositing User: Zahra Otroj
URI: http://eprints.mui.ac.ir/id/eprint/12280

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