Inovio Pharmaceuticals Inc (INO)

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(NEW)Optimized and enhanced DNA plasmid vector based in vivo construction of a neutralizing anti-HIV-1 envelope glycoprotein Fab

Kar Muthumani*, Seleeke Flingai, Megan Wise, Colleen Tingey, Kenneth E Ugen, David B Weiner
Kar Muthumani

*Corresponding author
Department of Pathology and Laboratory Medicine; University of Pennsylvania School of Medicine; Philadelphia, PA USA

Seleeke Flingai
Department of Pathology and Laboratory Medicine; University of Pennsylvania School of Medicine; Philadelphia, PA USA

Megan Wise
Department of Pathology and Laboratory Medicine; University of Pennsylvania School of Medicine

Colleen Tingey
Department of Pathology and Laboratory Medicine; University of Pennsylvania School of Medicine; Philadelphia, PA USA

Kenneth E Ugen
Department of Molecular Medicine; University of South Florida

Morsani College of Medicine; Tampa, FL USA; Center for Molecular Delivery; University of South; Tampa, FL USA

David B Weiner
Department of Pathology and Laboratory Medicine; University of Pennsylvania School of Medicine; Philadelphia, PA USA




Abstract
Monoclonal antibody preparations have demonstrated considerable clinical utility for the treatment of specific malignancies, as well as inflammatory and infectious diseases. Antibodies are conventionally delivered by passive administration, typically requiring costly large-scale laboratory development and production. Additional limitations include the necessity for repeat administrations, and the length of in vivo potency. Therefore, the development of methods to generate therapeutic antibodies and antibody like molecules in vivo, distinct from an active antigen-based immunization strategy, would have considerable clinical utility. In fact, adeno-associated viral (AAV) vector mediated delivery of immunoglobulin genes with subsequent generation of functional antibodies have recently been developed. As well, non-viral vector mediated nucleic acid based delivery technology could permit the generation of therapeutic/prophylactic antibodies in vivo, obviating potential safety issues associated with viral vectors. This delivery strategy has limitations as well, mainly due to very low in vivo production. In this report we have constructed an “enhanced and optimized” DNA plasmid technology to generate immunoglobulin heavy and light chains (i.e., Fab fragments) of an established neutralizing anti-HIV envelope glycoprotein monoclonal antibody (VRC01). The “enhanced” DNA (E-DNA) plasmid technology includes codon/RNA optimization, leader sequence utilization, as well as targeted potentiation of delivery and expression of the Fab immunoglobulin genes through use of adaptive in vivo electroporation. The results demonstrate that delivery by this method of a single administration of the optimized Fab expressing constructs resulted in generation of Fab molecules in mouse sera possessing high antigen binding and HIV neutralization activity for at least 7 d against diverse HIV isolates. Importantly, this delivery strategy resulted in a rapid increase (i.e., in as little as 48 h) in Fab levels when compared with protein-based immunization. The active generation of functional Fab molecules in vivo has important conceptual and practical advantages over conventional ex vivo generation, purification and passive delivery of biologically active antibodies. Further study of this technique for the rapid generation and delivery of immunoglobulin and immunoglobulin like molecules is highly relevant and timely.


https://www.landesbioscience.com/journals/vaccines/article/26498/