Abstract
Disialoganglioside GD2 is an important target on several pediatric and adult cancer types including neuroblastoma, retinoblastoma, melanoma, small-cell lung cancer, brain tumors, sarcomas, and cancer stem cells. We have utilized structural and computational methods to refine the framework of humanized monoclonal antibody 3F8, the highest affinity anti-GD2 antibody in clinical development. Two constructs (V3 and V5) were designed to enhance stability and minimize potential immunogenicity. Construct V3 contained 12 point mutations and had higher thermal stability and comparable affinity and in vitro tumor cells killing as the parental hu3F8. Construct V5 had nine point mutations to minimize potential immunogenicity, but resulted in weaker thermal stability, weaker antigen binding, and reduced tumor killing potency. When construct V3 was combined with the single point mutation HC:G54I, the resulting V3-Ile construct had enhanced stability, antigen binding, and a nearly sixfold increase in tumor cell killing. The resulting product is a lead candidate for clinical development for the treatment of GD2-positive tumors.
Highlights
GD2 is a ganglioside expressed in several pediatric and adult cancer types and has been actively targeted by cancer immunotherapy approaches
GD2 has been found to be expressed in neuroectoderm-derived tumors and sarcomas, including neuroblastoma, retinoblastoma, melanoma, small-cell lung cancer, brain tumors, and sarcomas [3,4,5]
Purified anti-GD2 monoclonal antibodies (MoAbs) were diluted in HBS-E buffer containing 250 mM NaCl at increasing concentrations (50–1600 nM) prior to analysis
Summary
GD2 is a ganglioside expressed in several pediatric and adult cancer types and has been actively targeted by cancer immunotherapy approaches (see Ref. [1] for recent review). GD2 is a member of the b-series gangliosides, which are normally expressed during fetal development and are highly restricted to the central nervous system in healthy adults, with low levels of expression on peripheral nerves and skin melanocytes [2]. GD2 has been found to be expressed in neuroectoderm-derived tumors and sarcomas, including neuroblastoma, retinoblastoma, melanoma, small-cell lung cancer, brain tumors, and sarcomas [3,4,5]. Because of its surface expression on tumor cells and restricted normal expression in the brain and low levels in the periphery, GD2 has been an ideal target for the development of monoclonal antibodies (MoAbs), which cannot cross the blood–brain barrier. Several anti-GD2 antibodies have been developed and tested in the clinic over the past 20 years, primarily in pediatric neuroblastoma patients. Murine 3F8 was more recently humanized (hu3F8) based on complementarity determining region (CDR) grafting [13], and is currently in Phase
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
