SBIR-STTR Award

This project is focused on the development of a novel transgenic avian production platform that will provide human therapeutics in a rapid, cost-effective fashion. Specifically, we intend to create transgenic Japanese quail (Coturnix Coturnix sp. japonica) that produce humanized monoclonal antibody (huMAb) within the albumen (white) portion of their laid eggs. The anticipated demand for therapeutic huMAbs will require a production capacity greater than what is currently available in the worldwide industry. The rapid onset of sexual maturity in quail will result in the production of huMAb in a fourth of the time required by other transgenic platforms (e.g., chickens, mammals and plants). The primary goal of Phase I is to produce huMAb in quail egg white in sufficient quantities for subsequent detailed characterization. This will be accomplished by first identifying DNA constructs that express huMAb in vitro in a cell specific manner. Transgenic G1 hens will be generated to produce huMAb in quantities sufficient for glycosylation and avidity analyses to be initiated in Phase II of the project. Phase II will conclude with the production of a transgenic flock capable of generating quantities of huMAb required for clinical trials. The development of quail as a bioreactor platform will decrease the time required to evaluate potential huMAb therapeutics other biologics

Thesaurus Terms:
biotherapeutic agent, egg food product, genetically modified animal, immunologic substance development /preparation, monoclonal antibody, quail, technology /technique development CD40 molecule, antiantibody, bioreactor, complementary DNA, immunoglobulin G, lysozyme biotechnology, cell line, enzyme linked immunosorbent assay, microinjection, transfection

This phase II SBIR project will test the feasibility of producing large amounts of human monoclonal antibody into transchromosomic avian egg white. We will use a full length ovomucoid genomic locus expression construct created and tested in phase I, to drive expression of the monoclonal's heavy and light chain cDNA. Using these constructs we produced transgenic hens that expressed human monoclonal antibody into their egg whites. This antibody was purified and analyzed for antigen binding and effector function. The results of this analysis support the conclusion that the transgenic produced antibody exhibits antigen binding and effector function that is comparable to the same antibody produced in a mammalian cell culture. This phase II project will also utilize an improved gene delivery system to generate fully transchromosomic avian hens. We have recently generated transchromosomal founders that exhibit germline transmission of a 60MB artificial chromosome. The offspring of these birds contain single copies of the artificial chromosome in every cell. This represents a breakthrough in avian transgenesis which to this point has been restricted by the limited payload capacity of retroviral-mediated methods. We will utilize this improved avian transgenisis technique to deliver the two ovomucoid-based human monoclonal antibody expression vectors developed in phase I to stage I embryos. Transgenic human monoclonal antibody from GO, G1 and G2 birds will be analyzed for expression level, purified and characterized for bioactivity and glycosylation. We anticipate that the G2 production flock will be able to supply sufficient human monoclonal antibody to proceed to clinical trials