Upstream
Introduction lectures
Biopharmaceutical
- partly produced using a cell (yeast, E.coli, animal cell, animal)
- complex molecule
- injectable
- expensive
- used for life threatening disease
Defined Undefined
➢ Chemical structure / composition is ➢ Critical product attributes unknown
known ○ Extensive testing in clinical
➢ Critical product attributes known trials
○ Product failures
○ mechanism of action
➢ Process not understood
○ side effects ○ Process fixed early in
➢ Understanding of the process development
○ predict product quality ○ Process for trials =process
for market
○ Suboptimal process based
on old technology
○ Reject good batches/delay in
batch approval
Both make the biopharmaceutical very
expensive
Most are undefined, mainly due to Post translational modifications (PTMs)
Mainly glycosylation, effects:
- activity
- immunogenicity
- in-vivo half life (clearance from body)
- stability (storage)
Production platforms
➢ Bacteria → small proteins,
peptides, non glycosylated and
easily refolded proteins
➢ Yeast → more complex proteins
with limited glycosylation
➢ Animal cells → complex proteins
with extensive PTMs
,Approval path
Preclinical Cells, Basic safety: dosages
guinea Investigational new drug application
pigs
Phase 1 20-50 Pharmacological actions,
humans Product safety & side effects
Phase 2 50-200 Effectiveness: optimal dosage,
patients application scheme etc.
Phase 3 Hundreds Final safety and effectiveness
patients Very expensive
pharmaceuticals are very expensive:
- whole process costs 2 billion euros
- only 1 in 10 passes all the trials, 1 needs to make up for the other failures
fails in phase 3 → disaster for the company
Fast track
Orphan drugs
- smal patient group
- live threatening diseases
- clear therapeutic of public health advantage (e.g. COVID)
Biosimilars
= exact copies of originals that have gone off patent
,Animal cell culture
➢ In vitro studies
○ Basic cell physiology
○ Toxicity
○ Drug candidates
○ Food components
➢ Cell therapy
○ CAR-T cell cancer therapy
➢ Tissue engineering
○ cartilage (kraakbeen) repair
○ cultured meat
➢ Pharmaceutical proteins
○ EPO = hormone that will increase red blood cell production
normally made in kidney, EPO used for patients with kidney failure
➢ Viral vaccines
➢ Gene therapy (lecture 9 molecular virology)
○ gene delivery by AAV virus
➢ Sponge derived pharmaceuticals
○ sponge = animal
○ defense mechanism = production of complex chemical molecules
○ sponge cells in bioreactor for production
Cell line development
Some cells can escape this Hayflick limit like cancer cells → better cell line
- Infinite life span
- Acceptable growth rate
- Low growth factor dependence
- Suspension growth
- aneuploid = having an abnormal number of chromosomes in a haploid set
Transformed cells obtained by
- Mutagens
- Viruses
- Oncogens
- Spontaneous
- Tumors
Examples
- CHO (Chinese Hamster Ovary): epithelium
- SF-21/SF9 Spodoptera frugiperda: Ovary
, Growth
Media
animals cells need complex media, certain ingredients can only be added up until a certain
amount, because otherwise it will precipitate → limited growth
E. coli grows much faster → in case of contamination, E. coli will outgrow the animal cells
Serum
= fluid and solvent component of blood which does not play a role in clotting
Functions Problems
Growth factors ●Infectious agents
Transferrin (Fe) (viruses, prions)
Lipids ●Variable composition
Insulin ●Expensive
Shear protection ●High protein content
Detoxification problems in DSP
because of the problems, almost not used anymore
if really needed, serum from the patient themself is used
Shear sensitivity
Cause: lack cell wall, cel size
Reactor design: mixing, gas transfer
Comparison
mammalian insect yeast bacteria
nutritional complex complex simple simple
media cost 10-100 10-100 <1 <1
(€/dm3)
Secretion Yes yes/lytic variable No
PTMs ++ + +/- --
DSP simple simple complex complex
Scale-up difficult difficult simple simple
Introduction lectures
Biopharmaceutical
- partly produced using a cell (yeast, E.coli, animal cell, animal)
- complex molecule
- injectable
- expensive
- used for life threatening disease
Defined Undefined
➢ Chemical structure / composition is ➢ Critical product attributes unknown
known ○ Extensive testing in clinical
➢ Critical product attributes known trials
○ Product failures
○ mechanism of action
➢ Process not understood
○ side effects ○ Process fixed early in
➢ Understanding of the process development
○ predict product quality ○ Process for trials =process
for market
○ Suboptimal process based
on old technology
○ Reject good batches/delay in
batch approval
Both make the biopharmaceutical very
expensive
Most are undefined, mainly due to Post translational modifications (PTMs)
Mainly glycosylation, effects:
- activity
- immunogenicity
- in-vivo half life (clearance from body)
- stability (storage)
Production platforms
➢ Bacteria → small proteins,
peptides, non glycosylated and
easily refolded proteins
➢ Yeast → more complex proteins
with limited glycosylation
➢ Animal cells → complex proteins
with extensive PTMs
,Approval path
Preclinical Cells, Basic safety: dosages
guinea Investigational new drug application
pigs
Phase 1 20-50 Pharmacological actions,
humans Product safety & side effects
Phase 2 50-200 Effectiveness: optimal dosage,
patients application scheme etc.
Phase 3 Hundreds Final safety and effectiveness
patients Very expensive
pharmaceuticals are very expensive:
- whole process costs 2 billion euros
- only 1 in 10 passes all the trials, 1 needs to make up for the other failures
fails in phase 3 → disaster for the company
Fast track
Orphan drugs
- smal patient group
- live threatening diseases
- clear therapeutic of public health advantage (e.g. COVID)
Biosimilars
= exact copies of originals that have gone off patent
,Animal cell culture
➢ In vitro studies
○ Basic cell physiology
○ Toxicity
○ Drug candidates
○ Food components
➢ Cell therapy
○ CAR-T cell cancer therapy
➢ Tissue engineering
○ cartilage (kraakbeen) repair
○ cultured meat
➢ Pharmaceutical proteins
○ EPO = hormone that will increase red blood cell production
normally made in kidney, EPO used for patients with kidney failure
➢ Viral vaccines
➢ Gene therapy (lecture 9 molecular virology)
○ gene delivery by AAV virus
➢ Sponge derived pharmaceuticals
○ sponge = animal
○ defense mechanism = production of complex chemical molecules
○ sponge cells in bioreactor for production
Cell line development
Some cells can escape this Hayflick limit like cancer cells → better cell line
- Infinite life span
- Acceptable growth rate
- Low growth factor dependence
- Suspension growth
- aneuploid = having an abnormal number of chromosomes in a haploid set
Transformed cells obtained by
- Mutagens
- Viruses
- Oncogens
- Spontaneous
- Tumors
Examples
- CHO (Chinese Hamster Ovary): epithelium
- SF-21/SF9 Spodoptera frugiperda: Ovary
, Growth
Media
animals cells need complex media, certain ingredients can only be added up until a certain
amount, because otherwise it will precipitate → limited growth
E. coli grows much faster → in case of contamination, E. coli will outgrow the animal cells
Serum
= fluid and solvent component of blood which does not play a role in clotting
Functions Problems
Growth factors ●Infectious agents
Transferrin (Fe) (viruses, prions)
Lipids ●Variable composition
Insulin ●Expensive
Shear protection ●High protein content
Detoxification problems in DSP
because of the problems, almost not used anymore
if really needed, serum from the patient themself is used
Shear sensitivity
Cause: lack cell wall, cel size
Reactor design: mixing, gas transfer
Comparison
mammalian insect yeast bacteria
nutritional complex complex simple simple
media cost 10-100 10-100 <1 <1
(€/dm3)
Secretion Yes yes/lytic variable No
PTMs ++ + +/- --
DSP simple simple complex complex
Scale-up difficult difficult simple simple
