BUSM 2428 Regulatory, ethical and legal issues in biotechnology
Study Period 3 2012
ChemTech Case study
Development of an avian flu vaccine for the Australian poultry industry
The company
ChemTech is a successful biotechnology company developing and manufacturing veterinary small molecule anti-infective drugs for production animals.
The company has decided to move into the development of recombinant vaccines for the treatment of respiratory diseases in intensively reared pigs and poultry. The company’s first project will be the development of a recombinant flu vaccine to treat avian influenza A (AIA) for the poultry industry in Australia. The company intends to develop the recombinant vaccine in house and outsource vaccine manufacture to Bioproperties Pty Ltd and field testing in a commercial hatchery (Baiada Poultry) under a collaborative research agreement with the Poultry Cooperative Research Centre Poultry Hub.
Company capabilities
ChemTech supports an active R&D section with expertise in chemical synthesis and analytical chemistry, microbiology and small molecule drug development and testing in vitro and in vivo. The company has laboratory facilities for chemistry, microbiology and cell culture and an animal facility capable of handling small animals and birds. It intends to establish capabilities for genetic engineering.
The AIA vaccine project
The company proposes to develop a human recombinant adenovirus serotype 5 (AD5) vectorencoding the haemagglutinin (HA) genes of pathogenic H5 or H7 AIA serotypes. The vaccine will be injected into chick eggs. The project has the following stages
In house laboratory studies (ChemTech)
1Development and in vitro evaluation of the vaccine construct
1.1Construction of replication-incompetent AD5 vector with H5 or H7 gene inserts
1.1.1PCR-amplification of H5 or H7 genes from a plasmid template using suitable primer pairs
1.1.2Insertion of the HA gene into the shuttle plasmid (pAdApt) to generate a plasmid with H5 or H7 gene (pH5/H7) and human CMV early promoter
1.2Construction of the recombinant Ad5 vector (Ad5-H5/H7)
1.2.1Co-transfection of human PER. C6 cells (Crucell) with the plasmid (pH5/H7) and the AdEasy™ adenoviral vector system.
1.2.2Separation of Ad5 vector clones by plaque assays
1.2.3 Validation of the construct by DNA sequencing
1.2.4Titration (ifu/mL) by the Adeno-X rapid titer kit
1.2.5stability & cell location of vector
2Antibody production in chicks
2.1 Vaccination by automated injection of single-doses of vaccine (106 to 1010 infectious units) into specific-pathogen-free (SPF) chicken eggsat day 18 of incubation (SPAFAS Australia Pty Ltd). SPF eggs meet European Pharmacopoeia SPF requirements.
2.2Evaluation of antibody response
2.2.1Determination of percent of chicks showing seroconversion at 21 and 42 days after hatch
HI inhibition or agar gel immunodiffusion (AGID) determination of antibody titres in serum samples from individual chickens against a pathogenic H5 or H7 strain.
2.2.2Determination of effective vaccination infection dose
2.3.2Characterisation of antibody binding (affinity, avidity, specificity & cross reactivity) to homologous AI strains using standard tests
3Demonstration of protective efficacy by AI challenge
3.1Challenge of cohorts of vaccinated and un-vaccinated SPF chicks hatched from bought-in SPC eggs (SPAFAS) with escalating doses (x times EID50) of a current AI strain with HA homology with the H5 or H7 vaccinating strain.
3.2Assessment of vaccine efficacy by
i) morbidity and mortality in vaccinated vs non-vaccinated chicks
ii) monitoring of AI infection and viral shedding in chicks by detection of viral RNA by AG precipitation & ELISA in cloacal samples
iii) antibody titres from serum samples
4Manufacture of the vaccine (Bioproperties)
Virus (Ad5-H5/H7)culture in human PER 6 cells in serum free suspension bioreactors and chromatographic purification
Formulation of vaccine doses for in ovo delivery
Commercial automated injector for eggs
5Field trial under commercial use conditions
Demonstration of clinical efficacy in broilers
Baiada Poultry commercial hatchery and broiler farm
5.11000 42 day chicken broiler study of vaccinated vs 1000 normally bred broilers under commercial hatchery conditions
5.2Assessment of vaccine efficacy as per 3.2
5.3Assessment of vaccine safety (Pharmacology)
5.4Assessment of environmental impact
Senior management at Chemtech have requested a summary report from the Project Manager on the regulatory requirements associated with the project work described above to assist them in planning execution of the project and to ensure the company is aware of all legal obligations relating to project studies undertaken in house and outsourced to vendors. Management wishes to be fully informed of its legal responsibilities as vaccine developer and contractor. This would include regulatory requirements relating to data submitted for product approval of the novel vaccine and licenses, permits and accreditations required by entities performing the listed studies. At this stage management does not require information about the pharmacological evaluation of the vaccine for safety (5.3) and possible environmental impact (5.4).