IIBBA   05544
INSTITUTO DE INVESTIGACIONES BIOQUIMICAS DE BUENOS AIRES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Selection of single domain antibodies against surface human melanoma antigens
Autor/es:
LACREU ML, ARRIAGA JM, BLANCO PA, BRAVO AI, MORDOH J, URRUTIA M; MORDOH J; URRUTIA MARIELA
Lugar:
Rosario
Reunión:
Conferencia; L SAIB; 2014
Institución organizadora:
SAIB
Resumen:
Cutaneous melanoma is a malignant tumor whose incidence and mortality rates have been increasing worldwide. Although new therapeutic options have improved therapy of advanced melanoma, the long-term prognosis remains poor. Therefore, the discovery of new targets is essential to design effective therapies. We have developed a phage display library constituted by variable domains of llama heavy-chain antibodies (VHH). VHHs possess small size, ability to recognize epitopes inaccessible to conventional antibodies, low immunogenicity and good tissue penetration. After enrichment of the library in VHHs that recognize melanoma antigens (Ags) through negative (using red blood cells) and positive (melanoma cells) steps, groups of phages-VHHs showed reactivity against melanoma patients tissue microarrays. Dry cell ELISA enabled us to identify 3 clones with strong binding against melanoma cells. Flow cytometry assays allowed us to select 4 high reactivity clones, one of them recognizing melanoma cells (MC) but not breast cancer cells (BC) or lymphocytes. The 3 others clones, which overlap those identified by ELISA, showed high reactivity to MC and BC. VHH clones with differential cell type recognition probably bind different antigens. The four binder clones were isolated and soluble VHHs produced. Binding assays of soluble VHHs and future perspectives for Ags identification will be presented.