Construction of VHH library derived from camelids

• Grant number: 23/07454-9
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: August 15, 2023
• End date: November 26, 2023
• Field of knowledge: Agronomical Sciences - Veterinary Medicine
• Principal Investigator: Débora Botéquio Moretti
• Grantee: Heloísa de Matos Trinca
• Supervisor: Gladys Viviana Parreño
• Host Institution: Instituto Butantan. São Paulo , SP, Brazil
• Institution abroad: Instituto Nacional de Tecnología Agropecuaria (INTA), Argentina
• Associated to the scholarship: 22/15549-7 - Evaluation of the immunotherapeutic potential of nanobodies against scorpion toxins, BP.MS

Abstract

Nanobodies (Nbs or VHHs) have great therapeutic potential and have been studied as a treatment for numerous diseases. In addition to having greater economic efficiency of production, these camelid-derived single-domain antibody fragments are small, stable, soluble in aqueous solutions, and have high specificity and affinity for the target antigen. In addition, VHHs exhibit low toxicity and immunogenicity due to their small size, identity to the human region of the heavy chain and rapid renal clearance. These characteristics and the ability to adapt nanobodies for in vivo applications make these molecules a promising and innovative generation of therapeutic proteins. The use of this technology is an underexplored strategy in Brazil. Due to the wide applicability of this tool, this work aims to amplify the VHH gene repertoire of llama species that may be used to build a naive library of camelid nanobodies. Peripheral lymphocytes will be isolated and quantified. The total RNA will be extracted and, subsequently, used for cDNA synthesis. The synthesized material will be submitted to sequential PCRs using specific oligonucleotide primers for the VHH. The amplified sequences of the VHH gene repertoire will, then, represent the variability of nanobodies of the animal and will be used for the construction of a library of VHH with the perspective of nanobody selection. (AU)