Nanobodies/antibodies to modulate and detect CXCR4 and CXCR7 mono-, homo-, and heteromers
Early stage researcher 3 (ESR3) project
Supervision: Bas van der Woning, Michael Saunders and Hans de Haard.
Host: arGEN-X (BE)
I- Project proposal:
1. Generate nanobodies/ SIMPLE antibodiesTM to specifically target the extracellular and intracellular site of CXCR4, CXCR7 mono-, homo-, and heteromers, in order to modulate their function or use as diagnostics.
2. Optimize the nanobodies/ SIMPLE Antibodies™ using ADCC-enhancing technology for in vivo modulation and diagnostics.
In order to identify nanobodies and antibodies against CXCR4 and CXCR7 llamas will be immunized with CXCR4 and CXCR7 encoding DNA and recombinant purified CXCR4 and CXCR7. Phage libraries will be generated displaying the variable domains of the antibody repertoire of the immunized llamas. Antigen specific clones will be selected by phage pannings on CXCR4/ CXCR7 bearing virus-like lipoparticles, and characterized by FACS analysis and cell-based assays. Full IgGs, optimized for inducing ADCC, will be produced for in vivo efficacy studies in mouse tumor models.
Planned secondment: VU (NL).
II- Requirement candidate:
Required diploma: MSc degree in molecular/biomedical Life Sciences, Pharmaceutical Sciences or related Life Science degree.
Required expertise: cell culture, cell-based assays, biochemistry and molecular biology. Recommended expertise: FACS analysis, recombinant DNA technology (PCR, restriction cloning, transformation, etc), phage display, protein purification, SDS-PAGE, working experience in industry.
1. Cuende J and Liénart S et al, and Lucas S (2015) Monoclonal antibodies against GARP/TGF-β1 complexes inhibit the immunosuppressive activity of human regulatory T cells in vivo. Science Translational Medicine. Accepted for publication.
2. Silence K et al., and De Haard HJ (2014) ARGX-110, a highly potent antibody targeting CD70, eliminates tumors via both enhanced ADCC and immune checkpoint blockade. MAbs 6:523-32.
3. Basilico C et al, and Michieli P (2014) Four individually druggable MET hotspots mediate HGF-driven tumor progression. J Clin Invest. 124:3172-86.