A fully human inhibitory monoclonal antibody to the Wnt receptor RYK.
RYK is an unusual member of the receptor tyrosine kinase (RTK) family that is classified as a putative pseudokinase. RYK regulates fundamental biological processes including cell differentiation, migration and target selection, axon outgrowth and pathfinding by transducing signals across the plasma membrane in response to the high affinity binding of Wnt family ligands to its extracellular Wnt inhibitory factor (WIF) domain. Here we report the generation and initial characterization of a fully human inhibitory monoclonal antibody to the human RYK WIF domain. From a naïve human single chain fragment variable (scFv) phage display library, we identified anti-RYK WIF domain-specific scFvs then screened for those that could compete with Wnt3a for binding. Production of a fully human IgG1κ from an inhibitory scFv yielded a monoclonal antibody that inhibits Wnt5a-responsive RYK function in a neurite outgrowth assay. This antibody will have immediate applications for modulating RYK function in a range of settings including development and adult homeostasis, with significant potential for therapeutic use in human pathologies.
Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia ; Angiogenesis Laboratory, Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Parkville, Victoria, Australia.
Michael M
MM
Halford
Maria L
ML
Macheda
Clare L
CL
Parish
Elena A
EA
Takano
Stephen
S
Fox
Daniel
D
Layton
Edouard
E
Nice
Steven A
SA
Stacker
D000818:Animals; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004285:Dogs; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D041681:NIH 3T3 Cells; D016501:Neurites / Q000276:immunology; D011518:Proto-Oncogene Proteins / Q000276:immunology; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology*; D051153:Wnt Proteins / Q000276:immunology; D000071818:Wnt-5a Protein
D057134:Antibodies, Neutralizing; D011518:Proto-Oncogene Proteins; D057127:Single-Chain Antibodies; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; C080836:RYK protein, human; D020794:Receptor Protein-Tyrosine Kinases