VLP_production_SOW.docx

STATEMENT OF WORK

cGMP VLP Production

1) BACKGROUND INFORMATION

The Laboratory of Infectious Diseases (LID) is dedicated to improving public health by studying viruses that cause human morbidity and mortality. Infections caused by influenza viruses are a significant health burden on the United States, causing up to 49,000 deaths per year (Centers for Disease Control). The current seasonal vaccine is produced each year with three-four influenza A and B virus strains predicted to circulate in the upcoming season. Although the seasonal influenza vaccine is a widely promoted health campaign, it is ill-suited to fight an emerging influenza pandemic or to offer protection against antigenically drifted epidemic strains. First, the efficacy of the vaccine depends on an accurate prediction of what strains will be in circulation because a close antigenic match between vaccine and circulating virus strain is required for protective efficacy. Additionally, the vaccine is usually grown in an egg-based system, which takes at least six months to produce vaccine stocks. If there were an unexpected emergence of a strain (i.e., a pandemic or a significant drift epidemic), production of a specific vaccine would not be ready in time. Accordingly, to protect against a pandemic, an alternative vaccine strategy should be pursued allowing broad protective efficacy and not requiring specific viral antigen matching. An ideal ‘universal’ influenza vaccine should be able to provide broad reactivity against drifted seasonal and pre-pandemic/pandemic viruses and be produced quickly to meet public health needs.

We have pursued a viral-like particle (VLP) platform to produce a practical ‘universal’ vaccine strategy. VLPs are non-infectious, genomeless particles that mimic the shape of actual viruses. This means that they do not cause disease, but can be recognized by the immune system in a similar way to the native virus. Additionally, VLPs can be mass-produced in a matter of weeks, as opposed to egg-based vaccines, which would take six months.

To generate a vaccine with broad protection, we have investigated the combination of different antigens from distinct influenza A virus hemagglutinin subtypes. Thus, we have previously purchased non-cGMP VLP antigens for pre-clinical animal experiments. To achieve our goal, these VLPs have been produced with the utmost quality and purity.

2) PURPOSE AND OBJECTIVES OF THE ACQUISITION

The goal is this project is to obtain cGMP baculovirus-produced VLPs for subsequent use in human clinical trial experiments after appropriate toxicity and sterility testing is performed. Vendor will use prior experience with influenza VLP and vaccine manufacturing to prepare cGMP certified batches of Quadrivalent Influenza VLP vaccine for subsequent use in these trials.

3) CONTRACTOR REQUIREMENTS

To produce a minimum of 5 mg of hemagglutinin (HA) protein in a purified cGMP influenza viral-like particle stock, the contractor will:

1) Obtain a pre-Master Cell Bank (pMCB, sometimes referred to as a Master Cell Seed) and will generate a Master Cell Bank (MCB)

a) If required by the FDA, a Working Cell Bank (WCB) will be subsequently generated.

2) MCB or WCB will be used in preparation of the Quadrivalent Influenza VLP vaccine for an IND Enabling Toxicology Study and Phase I Clinical Trial.

a) If required, pMCB and/or license rights for pMCB will be obtained by vendor from a current influenza vaccine manufacturer, reference storage facility, biopharmaceutical vendor, or other reputable source.

3) Provide pre-Master Virus Bank (pMVB, sometimes referred to as Master Virus Seed) for preparation of each of the four VLP subtypes required for manufacturing of Quadrivalent Influenza VLP vaccine.

a) Correspondingly, vendor will prepare four Master Virus Stocks (MVS).

b) If required by Client or the FDA, four Working Virus Stocks (WVS) will be subsequently prepared.

4) MVS or WVS will be used for preparation of the Quadrivalent Influenza VLP vaccine for an IND Enabling Toxicology Study and Phase I Clinical Trial.

5) The VLP batches will be manufactured in Vendors cleanroom facility that has been configured to produce Phase I clinical trial material according to the FDA 2008 Guidance for Industry “cGMP for Phase 1 Investigational Drugs”.

6) Each subtype VLP will be manufactured at the yield at least 5 mg of HA protein in Sf9 cells in Serum-Free Medium without antibiotics.

a) HA quantitation will be performed using single-radial immunodiffusion test (SRID) or other FDA acceptable method.

b) The purity, stability and suitability of the VLPs for the IND enabling GLP Toxicology and subsequent Phase I Clinical Trial are the shared responsibility of vendor and LID.

To purify VLPs to >90% purity, the contractor will employ one of the following techniques:

a) Employ tangential flow filtration and/or affinity chromatography

b) Purify VLPs using 2x chromatography

c) Run products through a 0.2 micron membrane filter Quality control:

a. Assessment of protein content (bicinchoninic acid assay (BCA) and/or SDS page)

b. Epitope confirmation (Western blot, electron microscopy)

c. Hemagglutination or Neuraminidase Inhibition assay

d. All steps in the expression and purification process will be performed in a manner that limits the accumulation of endotoxin. The final endotoxin specification target is 1 EU/mg protein. Endotoxin levels will be evaluated by established commercial methods by the contractor.

4) GOVERNMENT FURNISHED MATERIALS

· Plasmid stocks containing H1, H3, H5, H7 avian HA genes.

· Specific HA antibodies to detect HA in western blots or other detection assays.

5) CONTRACTOR DELIVERABLES

· Procure the required raw materials.

· Make available all in-house equipment.

· Generate and make available all required and appropriate documentation including, but not limited to, Experimental Procedures, Material Specifications, SOPs, etc.

· Perform the following in-process sampling and assays: - Sf9 Cell Density

- Sf9 Cell Viability - Baculovirus Plaque Assays - HA titers - SDS-PAGE - Western blots - Sterility - Mycoplasma assay

· Perform any additional in-process sampling and assays as requested by Client and not covered under this quotation or any other signed and relevant work order. - Additional sampling and assays will be executed at the expense of Client under a separate work order signed prior to the start and invoiced separately and directly to Client.

· Produce a cGMP MCB for Sf9 cells.

· Produce cGMP MVS for H1 VLP

· Produce cGMP MVS for H3 VLP

· Produce cGMP MVS for H5 VLP

· Produce cGMP MVS for H7 VLP

· Produce Process Development Run for H1 VLP

· Produce an Engineering Run for H1 VLP

· Produce cGMP H1 VLP

· Produce cGMP H3 VLP

· Produce cGMP H5 VLP

· Produce cGMP H7 VLP

· Produce Quadrivalent VLP formulation

· Provide the draft Batch Production Records to Client for review and approval prior to execution.

· Provide labeling for the VLP Batches.

· Provide a final QC report and copy of the completed Batch Production Record.

· Provide temporary 30 day storage of the materials.

6) INSPECTION AND ACCEPTANCE

· Documentation as required by FDA

· Bulk preparation of each VLP must pass industry standard FDA sterility and toxicity testing prior to final vial filling of quadrivalent vaccine cocktail

· Bulk preparation of each VLP must not contain infectious baculovirus

· A minimum of 5mg hemagglutinin (HA) protein of each VLP (H1, H3, H5, and H7) to be produced

· Final vial filling must pass subsequent sterility and toxicity testing

· Final vial filling with equal amounts of hemagglutinin (HA) protein of each of the four VLPs in the cocktail.

· Delivery of 80-85 final filled vials at a minimum concentration of 60ug of hemagglutinin (HA) protein from each of the four VLP hemagglutinins per 0.5ml in sterile saline at pH range 6.4-7.9, with a final volume of 0.5ml per each filled vial.