GENERATION OF NANOBODIES AS A BIOTECHNOLOGICAL TOOL FOR APPLICATION IN DIFFERENT AREAS OF BIOLOGY

ALZOGARAY V; FERNANDO A GOLDBAUM

Buenos Aires

Jornada; XXIV JORNADAS ANUALES DE LA SOCIEDAD ARGENTINA DE BIOLOGIA; 2022

Sociedad Argentina de Biología

Camelids naturally generate IgG antibodies formed only by heavy chain. The variable region of these antibodies that recognizes the antigen is called nanobodies (Nbs). These single domain antibody fragments have a series of properties among which stand out their small size, easy production, high specificity and affinity for the antigens they recognize, can penetrate cellular tissue, recognize hidden epitopes, are soluble and stable under critical conditions of temperature and pH. Through the application of different methodologies, Nbs can be obtained recombinantly and given their intrinsic properties are applied in areas such as diagnostics, therapeutics, imaging, immunotherapy, among others. Several Nbs are in phase I and II clinical trials, especially against cardiovascular, neurodegenerative and autoimmune diseases. In the present work we describe the pipeline we have put in place to obtain Nbs against different antigens which are mostly involved in diseases of local, regional and global relevance. The strategy includes the use of phage display technology to select Nbs that recognize the antigen under study. Subsequently, we employ the ELISA or nickel plate technique, which allows spatial orientation of the target protein, and as a result hundreds of Nbs can be characterized according to their specificity. The selected Nbs are expressed as recombinant protein and also characterized by their affinity. From this robust, versatile and simple platform we are able to obtain Nbs and use them as a biotechnological tool to apply them in different areas of biology such as electron microscopy, crystallographic studies, optogenetics studies and diagnostic assays.