Antibody Fragments to Improve HIV gp120 Protein Production

Camelid single domain antibody fragmentsVHHpossess a number of distinctive features including a small sizean elevated stabilityand an ability to recognize epitopes non accessible for conventional antibodiesBesides common research and immunotherapy applicationscamelid VHHs are also used as co crystallization chaperonesprotein aggregation controllers and enzyme activity tunersIn this proposalwe wish to utilize these unique VHH properties as a chaperone for HIV gpexpressionfolding and stability to improve gpprotein productionIn Phase Icamelid VHH antibodies to HIV gpwill be developed that are capable of enhancing gpexpression in yeast and in mammalian cells enabling a simple mode for rapid and scalable affinity purificationAt least afold improvement in gpproduction in mammalian cell transient expression will be the basis for phase II transitionThe effort in Phase II will be to develop stable VHH assisted gpprotein production CHO cell lines with significant improvement in yieldGMP manufacturing cell line development and Master Cell bank generation will be performedFollowing developmentscale up of upstream and downstream GMP processes will be completed