Victor Calvillo-Miranda

Worked under the supervision of Dr. Andrew Kruse in the Department of Biological Chemistry and Molecular Pharmacology (BCMP) -Discovered a novel nanobody targeting an integral membrane protein, muscarinic 3 acetylcholine receptor (M3R), of the G protein-coupled receptor (GPCR) family using a yeast display platform. -Optimized the recombinant expression and purification of directly labeled M3R from Sf9 insect cells for nanobody discovery campaign. -Biochemically characterized the M3R/nanobody interaction using radioligand binding assays, flow cytometry on mammalian cells, mutagenic screens, and pulldowns to profile its specificity among the muscarinic family and identify epitope. -Performed affinity maturation of M3R nanobody using a TWIST variant library on yeast display and characterized affinities through high-throughput recombinant nanobody expression and purification. -Currently exploring M3R-nanobody as a nanobody drug conjugate to increase potency and specificity of a previously characterized small molecule muscarinic receptor agonist. -Coauthored and collaborated on three publications spearheaded by other members of the Kruse lab to support science and communication in the lab spanning topics in GPCR biology, protein engineering, and nanobody discovery for therapeutically relevant targets. -Provided materials and advice to other academic researchers carrying out selections with different targets using the Kruse lab nanobody library platform.

Worked under the supervision of Dr. May Khanna. -Recombinant expression and protein purification of a target implicated in many human neurodegenerative diseases, TDP-43. -Small molecule library in silico docking against several TDP-43 constructs. -Biophysical binding assays of small molecule docking screen hits to identify TDP-43 binders using saturation transfer difference NMR, SPR, and microscale thermophoresis (MST). -In silico docking screen of small molecule library against novel RNA target, Malat1. -Established in vitro RNA transcription of Malat1 and purification protocol for the Khanna lab. -Small molecule binding assays of Malat1 docking hits using SPR and MST.