Production Technology for Recombinant Intravenous Immunoglobulin

DESCRIPTION provided by applicant Production Technology for Recombinant IVIg Organization GigaGen Inc PI David S Johnson Ph D The Specific Aim of this SBIR Phase I project is to develop a natural repertoire antibody protein expression system that will form the basis of a recombinant intravenous immunoglobulin rIVIg therapeutic product IVIg is a pool of proteins isolated from the sera of thousands of donors The FDA has approved IVIg therapy for six indications including idiopathic immune thrombocytopenic purpura ITP Kawasakiandapos s vasculitis B cell chronic lymphocytic leukemia CLL and primary immunodeficiencies Orange et al IVIg sales are $ billion worldwide and growing at per year due to an aging population and ever expanding off label modalities Taylor andamp Shapiro Unfortunately current methods for IVIg production threaten continued expansion of IVIg therapy because of supply chain risk impurities and contamination and batch to batch variation In this Phase I project we will take steps to demonstrate that we can express GigaMuneTM natural human repertoire DNA libraries a stable yeast expression system Analogous to Genentech years ago Russo our primary technology innovation is to use natural Ig repertoire DNA libraries expressed in a humanized Pichia yeast production system Li et al to replace a resource limited biological drug with a recombinant alternative To make rIVIg we will first use GigaGen GigaMuneTM technology to capture and re create expressed Ig repertoires from andgt blood donors GigaMuneTM uses advanced microfluidics and genomics to generate RT PCR libraries from millions of single cells per donor with native IgG subtypes and pairing between heavy and light chain We will then stably express the DNA repertoires en masse in engineered Pichia to produce massively polyclonal engineered rIVIg protein product Our rIVIg will have natural repertoire genetics engineerable content programmable glycosylation low production cost and consistent and predictable production We will accomplish the Specific Aim by performing the following tasks i Engineer a system for subcloning GigaLink tm DNA libraries en masse with native IgG isotype intact ii Optimize GigaLink tm DNA library delivery and stable display in a Pichia yeast production system and iii Use next generation sequencing NGS and Ig assays to assess several cell passages for uniformity and isotype content We will be successful if we achieve the following metrics i Use NGS to show that the Ig clone frequencies of the andgt diversity GigaLink tm libraries are maintained when subcloned and stably expressed in the yeast production system linear regression a power and ii Use NGS and antigen binding assays to demonstrate andlt CV between cell passages and time points one proportion z test a power Phase I will demonstrate that we can reproducibly produce high diversity GigaLink tm protein libraries in a yeast expression platform In Phase II we will take steps to build a GMP production facility and perform toxicology and pharmacokinetic studies on our rIVIg preparations At first rIVIg will simply substitute for conventional IVIg especially for patients who are deficient in antibodies i e hypogammaglobulinemia or humoral deficiencies Later our ability to engineer the content of the DNA library will open up broad new applications such as IgA deficiency and polyclonal anti tumor therapeutics PUBLIC HEALTH RELEVANCE Production Technology for Recombinant Intravenous Immunoglobulin Organization GigaGen Inc PI David S Johnson Ph D Intravenous immunoglobulin is used to treat many kinds of immune disorders and is currently derived from pools of blood from thousands of donors We are building new technology that will allow us to manufacture intravenous immunoglobulin without donor blood