IntroductionChikungunya virus (CHIKV) is a re-emerging mosquito transmitted alphavirus of global concern. Neutralizing antibodies and antibody Fc-effector functions have been shown to reduce CHIKV disease and infection in animals. However, the ability to improve the therapeutic activity of CHIKV-specific polyclonal IgG by enhancing Fc-effector functions through modulation of IgG subclass and glycoforms remains unknown. Here, we evaluated the protective efficacy of CHIKV-immune IgG enriched for binding to Fc-gamma receptor IIIa (FcγRIIIa) to select for IgG with enhanced Fc effector functions.MethodsTotal IgG was isolated from CHIKV-immune convalescent donors with and without additional purification by FcγRIIIa affinity chromatography. The enriched IgG was characterized in biophysical and biological assays and assessed for therapeutic efficacy during CHIKV infection in mice.ResultsFcγRIIIa-column purification enriched for afucosylated IgG glycoforms. In vitro characterization showed the enriched CHIKV-immune IgG had enhanced human FcγRIIIa and mouse FcγRIV affinity and FcγR-mediated effector function without reducing virus neutralization in cellular assays. When administered as post-exposure therapy in mice, CHIKV-immune IgG enriched in afucosylated glycoforms promoted reduction in viral load.DiscussionOur study provides evidence that, in mice, increasing Fc engagement of FcγRs on effector cells, by leveraging FcγRIIIa-affinity chromatography, enhanced the antiviral activity of CHIKV-immune IgG and reveals a path to produce more effective therapeutics against these and potentially other emerging viruses.

Zepteon: On demand antiviral antibody production

Author: Diamond, Michael ; Boesch, Austin ; Bolton, Glen ; Fox, Julie

Zepteon, Inc. is a privately held corporation founded in 2012 that has developed a first in class chromatog. resin, Glycap-3A, capable of separating antibodies with specific sugar structures (nonfucosylated) that are more potent against viruses and cancer.The affinity purification resin contains optimally immobilized FcgammaR3A receptors, leading to high antibody equilibrium binding capacities.Hyperimmune IgG is typically obtained from human donors who have been exposed to a virus and have generated virus specific antibodies.Literature indicates hyperimmune IgG would be far more potent against a range of viruses (Flu, MERS-CoV, Ebola, HPV, West Nile, CHIKV) using optimal IgG glycoforms and subclasses.In this talk, data showing that anti-viral plasma IgG fractionated using the Glycap-3A resin is far more potent against viral infection in mice will be presented.Zepteon resin is the only technol. capable of simultaneously providing more potent IgG glycoforms and subclasses (IgG1 and IgG3) for the $1B USD hyperimmune IgG market.The resin can be used to rapidly obtain therapies after a viral outbreak.The method is faster than the generation, testing and scale up of monoclonal antibody therapies.The technol. can enrich the most relevant IgGs from recently infected people to overcome viral strain mutation.In addition polyclonality provides multi-prong approach against viral escape.Zepteon is currently out-licensing the technol. to IVIG and hyperimmune manufacturers for up-front, licensing and milestone payments

Use of Fc gamma receptors as chromatographic ligands to enrich high affinity subclasses and glycoforms from human plasma IgG

Author: Kappel, Hunter ; Alter, Galit ; Ackerman, Margaret ; Boesch, Austin ; Mahan, Alison ; Chu, Ty

Human plasma is used as a manufacturing feedstock for a number of licensed IgG therapeutics in both immunity and infectious disease.Part of the mechanism of action for some of these therapies relies on the engagement of Fc gamma receptors (FcgRs) on immune cells, which in turn elicit effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis.Leveraging FcgRs as chromatog. affinity ligands, we were able to enrich in high affinity subclasses and glycoforms using human plasma derived IgG as a load material.These elution peaks were addnl. evaluated for their potentiation of both Natural Killer cell degranulation (ADCC surrogate) as well as Monocyte phagocytosis in cell-based assays.This work demonstrates the use of FcgR-based chromatog. to directly impact IgG effector function and illustrates the potential use of this technol. to develop new therapeutics from plasma IgG.

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