Vector and strain system for the in vivo conversion of antibody fragments into IgG molecules

Axiomx, Inc., Branford CT

Investigators

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Abstract

? DESCRIPTION (provided by applicant): Current methods for converting an antibody fragment (for example, scFv, Fab, Fab', etc.) involves subcloning from an M13 phagemid or yeast display vector by either restriction enzyme digestion or PCR, followed by ligation into 1 or more vectors to produce the heavy and light chains, transformation into E. coli and subsequent DNA sequencing validation. The proposed scFv --> IgG method using recombineering changes this paradigm for subcloning antibody fragments to produce IgG molecules into a simple transduction of a properly modified phagemid or yeast-display vector into a genetically-modified strain of E. coli harboring a specially-modified IgG expression shuttle plasmid. The costs and labor are reduced from approximately $100-$200 for traditional subcloning and DNA sequencing analysis to under $20 per clone conversion. Additionally, automation equipment is unnecessary and the low error-rate of recombineering is expected to obviate the need for DNA sequence validation. AxioMx is developing a pipeline for the rapid discovery (less than two weeks) of recombinant Abs. Completion of the objectives of this proposal will allow researchers to develop and quickly assemble IgG molecules which could be useful for high throughput proteome analysis, diagnostics, and immunotherapeutics. The ability to clone immuno-pools from phage, yeast and other display technologies while keeping the heavy and light chains linked is a significant advantage to the proposed method. We expect this method to increase the efficient production of better antibodies with implications for both diagnostics and therapeutics.