Lecture 4: Vaccine Dosage Form

Question 1

Smallpox

Answer 1

-Jenner and milkmaids
-cowpox immunes against smallpox

Question 1

Types of covid vaccines

Answer 1

-mRNA
-vector
-protein subunit
-whole, killed

Question 2

Polio vaccine

Answer 2

-whole vaccine

Question 3

Sterile suspensions of lipid nanoparticles (SLN)

Answer 3

-lipid shell that stabilizes mRNA vaccine
-Covid vax

Question 4

Covid mRNA vaccine

Answer 4

-mRNA in solid lipid nanoparticle

Question 5

Pfizer Vaccine

Answer 5

-DO NOT SHAKE
-energy from shaking can degrade vax nanoparticles

Question 6

mRNA vaccine (SLN)

Answer 6

-coated in lipid nanoparticles (endocytosed into cell)
-use mRNA to make viral spike protein
-immune system makes antibodies

Question 7

Vector Vaccines

Answer 7

-use another virus to carry in genetic info to make spike protein
-adenovirus for coronaviruses
-inject DNA that tells cells to make spike protein

Question 8

Protein Subunit Vaccine

Answer 8

-pieces of target virus
-infect moths to produce spike protein
-harvest spike protein

Question 9

Whole, killed vaccine

Answer 9

-killed with chemical or heat
-injected to illicit immune response

Question 10

RNA stability

Answer 10

-hydrolysis in blood or cells by basicity

Question 11

ionizable lipids

Answer 11

-positively charged at acidic pH
-neutral in blood
-ionization by henderson hasselbach equation

Question 12

Lipids in SLN

Answer 12

-cationic lipid
-helper lipids: cholesterol
-PEG lipid

Question 13

mRNA vax manufacture

Answer 13

-mix lipids in ethanol in solution 1
-mix nucleic acid dissolved in acidic buffer in solution 2

-2channels
-mix with syringe pump or flow system

Question 14

Manufacture lipid nanoparticles are

Answer 14

-densely packed lipids
-contain nucleic acids

Question 15

Other applications of SLNs

Answer 15

-maybe cancer and HIV
-theoretically could replace any protein in the body = tweak RNA code
-could encode growth factors

Question 16

Lipid nanoparticle structure

Answer 16

-coat mRNA in PEG lipid, ionizable lipid, phospholipid, cholesterol
-and nucleic acid

Question 17

CMOs

Answer 17

-contract manufacturing organizations
-effort to provide vax manufacturing services to companies

Question 18

RNA vax manufacturing workflow

Answer 18

1. Plasmid Linearization
2. In vitro transcription
3. Buffer exchange
4. Capture
5. Concentration
6. LNP formation
7. Polishing
8. Drug Product

Question 19

Making plasmid DNA

Answer 19

-development: selection and optimization
-quality and stability analysis
-cGMP (current goods manufacturing process) manufacturing

Question 20

1. Plasmid Linearization

Answer 20

-produce DNA template
-restriction enzyme opens circular DNA

Question 21

2. In vitro transcription

Answer 21

-synthesis of mRNA
-RNA polymerase3

Question 22

3. Buffer exchange

Answer 22

-prepare for purification

Question 23

4. capture

Answer 23

-purification of mRNA
-use resins
-remove small pieces of mRNA

Question 24

5. concentration

Answer 24

-titer

Question 25

6. LNP formation

Answer 25

-lipid nanoparticle formation
-mix with the mRNA
-particle size matters (DEATH)

Question 26

7. Polishing

Answer 26

-more purification

Question 27

8. Drug Product

Answer 27

-aseptic fill for drug product
-verify sterile products

Question 28

importance of solid lipid nanoparticles

Answer 28

-protect mRNA from hydrolysis in blood
-deliver mRNA to cell