● Design of plasmids for recombinant expression of monoclonal antibodies and antibody fragments● Generation of candidate molecules (mAbs and therapeutic enzymes) for biopharmaceutical development● Characterization of new anti-CD19 and anti-CD20 mAbs for immunotherapy via protein engineering techniques● Development and discovery of human monoclonal antibodies using soluble proteins and whole cells● High throughput analysis of mAb sequences by NGS (Illumina®)● Elaboration of kinetics assays (BLI, Octet®), nano-DSF and flow cytometry (staining and functional assays, apoptosis, ADCC and CDC) of monoclonal antibodyes (IgG) and their fragments (scFv and scFv-Fc)● Co-supervision and training of undergraduate and graduate students on protein engineering (mAbs and enzymes)● Expression and purification of recombinant proteins on prokaryotic (E. coli) and eukaryotic (Expi293 and ExpiCHO) platforms ● Establishment of stable pools (CHO) to improve protein yield, and development of clones for recombinant expression of monoclonal antibodies and antibody fragments● Operation, maintenance and training of instruments at the protein engineering platform (Octet® Red96e, Tycho nano-DSF, Akta Pure25, Celesta flow cytometer, Melody cell sorter)● Cell line culture for functional assay characterization of antibodies and antibody fragments, and therapeutic enzymes● Improvement of monoclonal antibody recombinant expression yield in >25-fold● Construction of a human antibody library for phage display campaigns of the group and collaborators

● Selection of monoclonal antibodies for immunotherapy using synthetic peptides and whole cells using phage display● Amplification of variable domains (heavy and light chain) for scFv library construction● Development of protocols for analyses of variable domain sequences obtained via phage display by next-generation sequencing (Illumina®)● FastQ analysis of antibody sequences by ATTILA (AuTomated Tool for Immunoglobulin Analysis) to propose enriched candidates from phage display campaigns● Recombinant expression of scFv candidate molecules in prokaryotic platform (Escherichia coli)● Implementation of the eukaryotic recombinant platform (ExpiCHO and Expi293)● Implementation and establishment of protocols for purification of antibodies (IgG1) and antibody fragments (scFv and scFv-Fc) by affinity and size exclusion chromatography using semiautomated system (Akta Start and Akta Pure25)● Establishment of protocols for antibody characterization by flow cytometry (staining, apoptosis, ADCC)● Implementation of the platform for protein-protein kinetics characterization using biolayer interferometry (BLI, Octet®)● Development of one anti-CD19 and two anti-CD20 monoclonal antibodies with kinetics (affinity) and functional (staining, apoptosis) features tuned by protein engineering techniques● Discussion groups and construction of pipelines combining structural bioinformatics and protein engineering● Evaluation of variants and monoclonal antibody clones by nano-DSF, ELISA, BLI and flow cytometry

● Directed evolution, via phage display, of one therapeutic monoclonal antibody● Obtention of five anti-CD20 scFv candidates● Implementation of the phage display and prokaryotic recombinant expression platforms● Establishment of protocols for phage display selection of antibodies● PCR, cloning, sequencing, recombinant expression and characterization of functional variants of one therapeutic antibody developed by directed evolution● Recombinant expression yield improvement in prokaryotic platform of scFv molecules from <1mg/L to >4mg/L

● Project and grant writing● Establishment of three molecular diagnostic tests (PCR) for animal health● Co-approval of two grants (R$230,000.00) for the molecular biology lab structuring● Formal project presentations to funding agencies

● Obtention of protein mutants via site-directed mutagenesis● Functional characterization of mutants by in vitro assays (actin polymerization assay) and mass spectrometry (protein disorder analysis)● Actin quantification of mutant cell lines by flow cytometry and confocal microscopy● Functional characterization of anti-CD20 antibody mutants by flow cytometry● Co-supervision of master student on their thesis “Molecular Characterisation of WASp Patient Mutations to understand Actin Regulation in T cells”, Master of Science da University of Applied Sciences (FH Campus Wien, Austria), with distinction

● Establishment of protocols for phage display library constructions for directed evolution● PCR, cloning and recombinant expression of antibody fragments● Establishment of protocols for epPCR library construction and diversity analysis of repertoire via Sanger and NGS (Illumina®)● Obtention of one library of mutants aiming directed evolution and affinity maturation of one therapeutic antibody

● Development of automation capable protocols for cell assays and bioprocess characterization (>99% reproducibility)● Optimization of protocols for cell detachment from microbeads, counting/viability and flow cytometry (cell cycle)

● Clinical and molecular profile of MEN1 patients● qPCR and genotyping