Modular Protein Manufacturing Platform

This request is based on successful conclusion of the phase one stage of developing a protein manufacturing platform implementing a novel vector system that can rapidly and efficiently generate cell lines capable of producing proteins at high yield and purity in a short time period. Lentigen is able to perform such a quick protein manufacturing process based on its LentiMax™ lentiviral (LV) gene delivery technology that stably delivers and permanently integrates sufficient number of copies of a gene to efficiently overexpress the encoded protein, and does it with up to 100 % efficiency in the target cell population. Lentigen is able to generate protein-producing mammalian cell lines quickly, in as little as four (4) weeks, because its LentiMax system is inherently flexible and modular allowing all nucleic acid sequences under 6 Kb in length to be rapidly cloned. Transduction of cells with lentiviral particles is simple and cell lines theoretically should be able to produce protein indefinitely. Lentigen has monitored one cell line and has demonstrated that is has produced protein for over a year and 64 generations of cell cultures without any positive selection of the cells and still able to recover the original high producer clones from a raw bulk cell culture. To date, as part of phase I study, Lentigen was able produce 15-28 gram per Liter of erythropoietin from cells that have been transduced with LentiMax. Lentigen also demonstrated that its technology allows the production of more complex proteins such as recombinant immunoglobulins as well and successfully implemented each step that is required to replace hybridomas for specialized high-yield protein producing cell lines: (a) isolating, (b) sequencing (c) optimizing and (d) rebuilding selected immunoglobulin genes in LentiMax constructs; (e) effectively transducing protein producing cell lines, (f) overexpressing the recombinant Ig genes at high per cell copy numbers (20-30 copies per cell), (g) producing functionally intact immunoglobulin molecules and optimizing the production conditions to manufacture at minimum ~0.5-1 g/L of recombinant murine immunoglobulins. Lentigen is confident that its LentiMax technology will be able to produce clonal cells that express secreted immunoglobulins of interest in amounts comparable to erythropoietin. With such results Lentigen proposes to build a new specialized platform technology optimized for Recombinant Ig Production (RIPP) which is essentially a lentivector based protein production plug-in system. Such an accomplishment would dramatically reduce the time and cost required to produce valuable proteins such as monoclonal antibodies both for diagnostic and clinical purposes at large scales. Lentigen is applying for this STTR to adapt its proven protein manufacturing technology to be able to produce proteins in its specialized proprietary CHO-derived cell lines in serum free medium, optimized for large scale clinical and commercial applications. The expectation would be to generate several grams of protein per liter in as little as 4 weeks from the identification the pathogen, and achieve commercial scale production within 8-12 weeks.