Lecture 4: Vaccine Dosage Form Flashcards
Smallpox
-Jenner and milkmaids
-cowpox immunes against smallpox
Types of covid vaccines
-mRNA
-vector
-protein subunit
-whole, killed
Polio vaccine
-whole vaccine
Sterile suspensions of lipid nanoparticles (SLN)
-lipid shell that stabilizes mRNA vaccine
-Covid vax
Covid mRNA vaccine
-mRNA in solid lipid nanoparticle
Pfizer Vaccine
-DO NOT SHAKE
-energy from shaking can degrade vax nanoparticles
mRNA vaccine (SLN)
-coated in lipid nanoparticles (endocytosed into cell)
-use mRNA to make viral spike protein
-immune system makes antibodies
Vector Vaccines
-use another virus to carry in genetic info to make spike protein
-adenovirus for coronaviruses
-inject DNA that tells cells to make spike protein
Protein Subunit Vaccine
-pieces of target virus
-infect moths to produce spike protein
-harvest spike protein
Whole, killed vaccine
-killed with chemical or heat
-injected to illicit immune response
RNA stability
-hydrolysis in blood or cells by basicity
ionizable lipids
-positively charged at acidic pH
-neutral in blood
-ionization by henderson hasselbach equation
Lipids in SLN
-cationic lipid
-helper lipids: cholesterol
-PEG lipid
mRNA vax manufacture
-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
Manufacture lipid nanoparticles are
-densely packed lipids
-contain nucleic acids
Other applications of SLNs
-maybe cancer and HIV
-theoretically could replace any protein in the body = tweak RNA code
-could encode growth factors
Lipid nanoparticle structure
-coat mRNA in PEG lipid, ionizable lipid, phospholipid, cholesterol
-and nucleic acid
CMOs
-contract manufacturing organizations
-effort to provide vax manufacturing services to companies
RNA vax manufacturing workflow
- Plasmid Linearization
- In vitro transcription
- Buffer exchange
- Capture
- Concentration
- LNP formation
- Polishing
- Drug Product
Making plasmid DNA
-development: selection and optimization
-quality and stability analysis
-cGMP (current goods manufacturing process) manufacturing
- Plasmid Linearization
-produce DNA template
-restriction enzyme opens circular DNA
- In vitro transcription
-synthesis of mRNA
-RNA polymerase3
- Buffer exchange
-prepare for purification
- capture
-purification of mRNA
-use resins
-remove small pieces of mRNA
- concentration
-titer
- LNP formation
-lipid nanoparticle formation
-mix with the mRNA
-particle size matters (DEATH)
- Polishing
-more purification
- Drug Product
-aseptic fill for drug product
-verify sterile products
importance of solid lipid nanoparticles
-protect mRNA from hydrolysis in blood
-deliver mRNA to cell
