CFI Pathogen Inactivation of Human Plasma Units

PROJECT SUMMARY
The rapid spread of the Zika virus, which can have a significant impact on neurological disorders in unborn
fetuses and potentially adults, the recent outbreak of the extremely virulent Ebola virus, periodic emergence of
SARS, recurrent outbreaks of potentially pandemic strains of influenza such as H5N1, the continuing epidemic
of MERS and the worldwide AIDS epidemic have highlighted a persistent concern in the health-care
community -- the need for effective pathogen inactivation and removal techniques for human blood plasma and
plasma-derived products. There is no commercially available, FDA-approved technology for the inactivation of
non-enveloped viruses in pooled human plasma and biologics, and only one approved method for units of
plasma, which can inactivate some, but not all known non-enveloped viruses. This dearth of FDA-approved
pathogen inactivation technologies could pose a significant future threat for known and new viruses in human
plasma and biologics. We propose to develop a physical pathogen inactivation technology, CFI™, for the
inactivation of both non-enveloped and enveloped viruses as well as pathogenic bacteria and parasites in
human plasma, plasma protein products and biologics. CFI™ technology is applicable to both pooled human
plasma and units of plasma, the more globally significant focus of the current application.
CFI™ (critical fluid inactivation) utilizes supercritical and near-critical fluids (SuperFluids™ or SFS).
SuperFluids™ are normally gases which, when compressed, exhibit enhanced thermodynamic properties of
solvation, penetration, selection and expansion. These gases are used to permeate and saturate virus and
pathogen particles. The SFS-saturated particles then undergo decompression and, as a result of rapid phase
conversion, virus inflation and rupture at their weakest points. We have demonstrated that the CFI™ (critical
fluid inactivation) process inactivates both enveloped viruses such as MuLV, VSV, Sindbis, HIV (all completely
inactivated), TGE, and BDVD, and the non-enveloped viruses Polio, Adeno, EMC (complete inactivation), Reo,
and Parvo viruses, while preserving biological activity of the CFI-treated product. In research collaboration
with the National Institute of Biological Standards and Control (NIBSC), London, England, our CFI inactivated
more than 4 logs of human Parvovirus B19 (one of the smallest and toughest viruses) in human plasma in a
two-stage CFI™ unit in less than 20 seconds. We have also demonstrated that SFS can disrupt and inactivate
microorganisms such as E. coli, thick-walled prokaryotes such as Bacillus subtilis and tough eukaryotes such
as Saccharomyces cerevisiae at viral inactivation SFS conditions. CFI can be used with viral reduction
methods such as nanofiltration as an orthogonal method of pathogen clearance, and is versatile for refinement
to treat cellular blood. The present data have been generated using prototypes of our pilot-scale CFI unit.
Our Phase I Specific Aims are to: (1) Design and construct a portable and comparably versatile bench-top CFI
unit for parallel treatment of single units of human plasma; and (2) CFI treat human plasma in the designed
bench-top device using a customized blood bag, and rigorously characterize the CFI-treated units of human
plasma for protein and enzyme activities. Our Phase II Specific Aims are to: (3) Complete characterization of
CFI-treated units of human plasma using customized blood bag in the bench-top CFI prototype; (4) Conduct
toxicological and neoantigenic studies of CFI treated human plasma in small animal models; and select best
CFI design in terms of performance, operations and cost; and (5) Draft IND, establish and conduct pre-IND
meeting with FDA, prepare and file a Drug Master File (DMF) for the manufacturing of CFI treated human
plasma units. In Phase III, we will construct bench-top CFI units for blood banks, and through licensing
agreements, provide equipment and technology transfer as well as prevention and maintenance support to
blood banks.PROJECT NARRATIVE
There are a number of emerging viruses such as Zika, West Nile, Ebola, SARS, potential pandemic strains of
influenza (H5N1), the Mexican swine flu, bacteria, parasites and a number of potential bioterrorism pathogens
such as smallpox that are of concern to the safety of the human plasma supply. Current approaches for
pathogen inactivation in biologics are not always effective against a wide spectrum of human and animal
viruses, are sometimes encumbered by process-specific deficiencies, and often result in denaturation of the
biologicals that they are designed to protect. CFI pathogen inactivation technology gives pathogens the
“bends,” inactivating them without damaging proteins and enzymes in medically important transfusion fluids
such as human plasma. This purely physical technique does not involve the use of heat, chemicals and/or
irradiation, each of which has significant drawbacks in the viral inactivation of human plasma. As such, while
CFI is capable of inactivating wide classes of viruses, bacteria and parasites, it has negligible negative impact
on biological integrity and potency of the treated fluids. We have developed, tested and validated this process
using continuous-flow laminar flow devices, and now plan to scale this technology down for units of human
plasma using a novel, portable bench-top CFI device using blood bags. The potential impact of a generally-
applicable physical technology for inactivating viruses and emerging pathogens with high retention of biological
activity will be very significant. CFI technology will also be impactful in developing countries and hot zones for
the clearance of viruses from human plasma.