Chapter 5 Vaccine Manufacturing

Question 1

What are the four (4) main competencies required of vaccine manufacturers?

Answer 1

1. Clear processes that define how the product is made
2. Compliance of the manufacturer to successfully complete these processes
3. Testing of the products and supporting operations
4. Regulatory authorization to release and distribute the product

Question 2

What are the four (4) main components of the process of vaccine approval?

Answer 2

1. Preparation of pre-clinical material for proof-of-concept studies
2. Submission of an investigational new drug application to the regulatory body
3. Testing for safety, immunogenicity, efficacy and effectiveness through clinical and non-clinical studies
4. Submission of all data (clinical, non-clinical, manufacturing) to the regulatory body for final review and licensure

Question 3

What are the five (5) main steps of traditional vaccine manufacturing (excluding mRNA vaccines)?

Answer 3

1. Cell culture/fermentation
2. Isolation
3. Purification
4. Formulation (including adjuvants)
5. Control and monitoring of operations

Question 4

Describe lyophilization.

Answer 4

Low temperature dehydration process, where the sample is first frozen, then brought to low pressure where the ice then sublimates.

Question 5

Describe the cell culture fermentation process for the anthrax vaccine BioThraz.

Answer 5

Growth of a microaerophilic culture of avirulent, nonencapsulated Bacillus anthracis in a chemically defined protein-free medium.

Question 6

Describe the cell culture fermentation process for the Haemophilus influenzae vaccine ActHIB.

Answer 6

Growth of H. influenzae type b strain 1482 in a semi-synthetic medium.

Question 7

Describe the cell culture fermentation process for the hepatitis A vaccine Havrix.

Answer 7

Propagation of Hep A strain HM175 in MRC-5 human diploid cells.

Question 8

Describe the cell culture fermentation process for the hepatitis B vaccine Recombivax HB

Answer 8

Production of recombinant Hep B surface antigen in yeast cells grown in a complex medium.

Question 9

Describe the cell culture fermentation process for the influenza vaccine Fluzone.

Answer 9

Propagation of the influenza virus on embryonated chicken eggs.

Question 10

Describe the cell culture fermentation process for the Japanese encephalitis vaccine JE-VAX.

Answer 10

Intracerebral inoculation of the virus in mice.

Question 11

Describe the cell culture fermentation process for the MMR vaccine.

Answer 11

Propagation of the measles virus and mumps virus in chick embryo cell culture and propagation of the rubella virus in WI-38 human diploid lung fibroblasts.

Question 12

Describe the cell culture fermentation process for the meningitis vaccine Menactra (A, C, Y, W-135).

Answer 12

Culture of meningococcal strains on Mueller-Hinton agar and growth in Watson-Scherp medium, and culture of Corynebacterium diphtheriae on modified Mueller and Miller medium for the toxoid component.

Question 13

Describe the cell culture fermentation process for the pneumonococcal vaccine Prevnar.

Answer 13

Growth of Streptococcus pneumoniae strains 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F on soy peptone broth, and growth of Corynebacterium diphtheriae in casamino acids for the CRM197 protein for the conjugate component.

Question 14

Describe the cell culture fermentation process for the polio vaccine IPOL.

Answer 14

Growth of poliovirus types 1, 2 and 3 in Vero cells on microcarriers using Eagle MEM modified meidum supplemented with newborn calf serum.

Question 15

Describe the cell culture fermentation process for the rabies vaccine RabAvert.

Answer 15

Growth of rabies virus in primary culture of chicken fibroblasts in a synthetic cell culture medium.

Question 16

Describe the cell culture fermentation process for the typhoid fever vaccine Vivotif.

Answer 16

Fermentation using a medium containing digest yeast extracta and acid digest of casein, dextrose and galactose.

Question 17

Describe the cell culture fermentation process for the yellow fever vaccine YF-Vax.

Answer 17

Culture of the yellow fever virus strain 17D-204 on living avian leukosis virus-free chicken embryos.

Question 18

Name six (6) methods for antigen isolation in vaccine manufacturing.

Answer 18

1. Centrifugation
2. Clarification
3. Release following cell disruption
4. Filtration
5. Homogeneization
6. Tissue harvest

Question 19

Name six (6) methods of antigen purification used in vaccine manufacturing.

Answer 19

1. Ultrafiltration and/or diafiltration
2. Phenol extraction
3. Chromatography
4. Flow centrifugation on linear sucrose gradient density
5. Centrifugation and supernatant collection
6. Chemical precipitation

Question 20

Name eight (8) adjuvants used in vaccine manufacturing.

Answer 20

1. Aluminum hydroxyphosphate sulfate (AAHS)
2. Aluminum hydroxide
3. Aluminum phosphate
4. Potassium aluminum phosphate
5. AS04 (monophosphoryl lipid A and aluminum salt)
6. MF59 (oil-in-water emulsion composed of squalene)
7. AS01_B (monophosphoryl lipid A and compound QS-21, extracted from Chilean soapbark tree)
8. CpG 1018 (cytosine phosphoguanine)

Question 21

Describe the typical strain content of seasonal influenza vaccine.

Answer 21

Two type A strains (H1N1 and H3N2) and one type B strain.

Question 22

Name two (2) major advantages of recombinant vaccines.

Answer 22

1. No potential for infection
2. Free of human blood products