CPS-322 Pharmaceutical Biotechnology // Most important bullet points
1. Introduction
1.1 Definitions
Biotechnology = Use of biological processes, living cells or organisms, to make products
useful for mankind. Example is beer. Often colour codes are used.
green = agricultural / white = industrial / red = medical / blue = aquatic
Pharmaceutical = Use of biological processes, living cells or organisms, to produce
Biotechnology therapeutic or diagnostic products. Not only medicine.
Biologic = Medicinal product extracted from a biological source; for example,
extracted insulin from pigs.
Biotechnology = Any pharmaceutical product used for therapeutic or in vivo diagnostic
Medicine purposes, which is produced in full or in part by either traditional or modern
biotechnological means
Biopharmaceutical = a protein or nucleic acid based pharmaceutical substance used for
therapeutic or in vivo diagnostic purposes, which is produced by means other
than direct extraction from a native (non-engineered) biological source.
Biological Medical Products.
Protein structures
Primary = the sequence of amino acids and residues
Secondary = recurring structural patterns (alpha helix, beta sheet)
Tertiary = 3D folding
Quaternary = spatial arrangement of protein subunits, more proteins together
Glycosylated = carbohydrate chains attached to protein backbone
1.2 History of biopharmaceuticals
Biologics (19th century) → Where it all began. Extracting medicinal products from biological sources
- Antiserum
o Tetanus antitoxin
o Diphtheria antitoxin (polyclonal antibodies from horse serum)
- Hormones/growth factors
o Insulin (1921, from pigs)
o Glucagon, FSH, Urokinase
There were problems.
- Immune reactions, contamination with infectious agents, short supply and batch-to-
batch variability
Recombinant DNA technology
Recombinant DNA technology boosted drug development in 1970s-1980s. Nowadays it is one of the
main production methods for biopharmaceuticals
Concept of recombinant DNA technology
1. mRNA strand which expresses gene of interest is isolated and cDNA is amplified using PCR
2. The cDNA sequence is transfected into expression vector
3. The expression host cells are upscaled leading to higher production of desired protein
(Upstream processing)
4. The desired proteins are recovered from the host cells and purified (Downstream
processing)
,1.3 SMD vs. BP
Small drug molecule Biopharmaceutical
+ Large diversity of chemical molecules + Any protein or nucleic acid can be used as drug
+ Well established technology + Quick discovery process
+ Cheap to manufacture + High specificity (less toxicity issues)
+ Control over chemical process is easy + Long half-life
+ Potential to cure diseases instead of alleviating
symptoms
- Discovery process is slow - Species specific
- Side effects & toxicity are common - Not orally active, don’t cross BBB, low
- Rapid elimination distribution & bioavailability
- Oral route not always an option - Longer development paths
- Potential distribution throughout the body - Expensive production
- Unstable & sensitive to heat/pH/etc.
- Control over biological process is difficult
2. Description & Registration
2.1 Classes of Biopharmaceuticals
Four major classes:
1. Extracted from living systems (animals)
a. Antibodies
b. Organs
2. Produced by recombinant DNA
a. Hormones
b. Coagulation factors & Antithrombotic factors
c. Growth factors & cytokines (interferons, interleukins)
d. (Therapeutic) monoclonal antibodies
e. Therapeutic enzymes
3. Vaccines
4. Gene therapy
a. Nucleic-acid based drugs (antisense oligonucleotides)
Hormones are used to communicate between organs and tissues for physiological regulation and
behavioural activities. They bind to specific receptor proteins in the target cell, which leads to an
activation signal transduction pathway. Commonly used hormones are oestrogens, progesterone,
and insulin.
Cytokines are a broad class of small proteins (5-20 kDa) and they play an important role in cell
signalling. They are molecules used for communication between cells to trigger the immune system
to remove pathogens. Cytokine pathways are an important target in a wide variety of diseases (auto
immune). They can be produced in vectors that are suitable for cloning (e.g. yeast cells, mammalian
cells or bacteria) when transfected with the desired gene(s), or can be modified by alteration of the
glycosylation pattern or changes in the amino acid sequence. Examples are
- Interferons
o Group of signalling proteins released in response to pathogens.
o Protect cells from virus infection
▪ Interferon beta-1a & beta-1b used to treat and control MS
▪ Can also be used to treat forms of cancer
- Interleukins
o Group of cytokines produced by white blood cells. Promote the development
and differentiation of T- and B-lymphocytes.
, ▪ Denileukin diftitox (Ontak) for cutaneous T-cell lymphoma
• Binds to an Interleukin-2 receptor and then releases diphtheria
toxin
Growth factors stimulate cell growth and proliferation in eukaryotic cells.
- Insulin-like growth factor (IGF-1)
o Polypeptide hormone similar to insulin
o Used to treat growth failure in children having trouble synthesizing IGF-1
Clotting factors are plasma glycoproteins that play a role in the blood coagulation pathway. Part of
complex chemical cascades at the site of bleeding which lead to blood clotting and finally
haemostasis.
Recombinant clotting factors are Factor VIII → Shortage leads to haemophilia A
Factor IX → shortage leads to haemophilia B
Recombinant clotting factors have an identical working mechanism to natural clotting factors:
1. The enzyme Thrombin activates recombinant Factor VIII cutting out its B-domain
2. Active recombinant Factor VIII interacts with co-factor IX and collectively activate FX
which indirectly leads to clot forming
3. Lack of either FVIII or FIX leads to excess bleeding like in haemophilia.
Enzymes catalyse chemical changes without being changed itself. They are made up of proteins.
Vaccines contain a pathogen (or a part of a pathogen) which is mostly weakened or killed. The
immune system responds and in case of next contact with the pathogen, the immune system
‘knows’ how to react and responds very quickly.
1. APCs take up and break down the antigen
2. APCs bind the peptide fragment to MHC molecule
3. APCs present the MHC antigen complex to T cells
4. T cells activate B cells
5. B cells produce antibodies
, Killed pathogens can still provoke an immune response because the immune system recognizes
PAMPs (pathogen associated molecular patterns) = molecules associated with groups of pathogens
that are recognized by cells of the innate immune system. Examples:
- Glycans, Glycoconjugates, Flagellin
- dsRNA, Bacterial lipopolysaccharides
They are recognized by Toll-Like receptors
Inactivated Vaccines
Inactivated vaccines use a killed version of the pathogen.
- Safe
- Might need several doses overtime to provide protection, because they usually don’t
provide immunity as strong as live vaccines
Live-attenuated vaccines
Live vaccines use a weakened (attenuated) version of the pathogen.
- Similar to natural infection → strong and long-lasting immune response
- 1 or 2 doses can give lifetime protection
- For people with weakened immune system, this can be dangerous
- Need to be kept cool
mRNA vaccines
Vaccines make (spike) proteins which trigger an immune response.
- Do not contain virus, so impossible to infect patient
- Short manufacturing times
Subunit, Recombinant, polysaccharide and conjugate vaccines
These vaccines use specific pieces of pathogen like its protein, sugar, or capsid.
- Give strong immune response to key parts of pathogen
- Safe for everyone (including patients with weakened immune systems)
- May need booster shots
Toxoid vaccines
Toxoid vaccines use a weakened/killed toxin made by the pathogen.
- Create immunity to the parts of a pathogen that creates the disease, not the pathogen
itself.
- Immune response is targeted to the toxin instead of the whole pathogen
- May need booster shots
Viral vector vaccines
Viral vector vaccines use a modified version of a different virus as a vector to deliver protection.
- Adenovirus is one of the most common vectors
- Delivers genetic info for the cell to make protein of the pathogen (like mRNA vaccines)
1. Introduction
1.1 Definitions
Biotechnology = Use of biological processes, living cells or organisms, to make products
useful for mankind. Example is beer. Often colour codes are used.
green = agricultural / white = industrial / red = medical / blue = aquatic
Pharmaceutical = Use of biological processes, living cells or organisms, to produce
Biotechnology therapeutic or diagnostic products. Not only medicine.
Biologic = Medicinal product extracted from a biological source; for example,
extracted insulin from pigs.
Biotechnology = Any pharmaceutical product used for therapeutic or in vivo diagnostic
Medicine purposes, which is produced in full or in part by either traditional or modern
biotechnological means
Biopharmaceutical = a protein or nucleic acid based pharmaceutical substance used for
therapeutic or in vivo diagnostic purposes, which is produced by means other
than direct extraction from a native (non-engineered) biological source.
Biological Medical Products.
Protein structures
Primary = the sequence of amino acids and residues
Secondary = recurring structural patterns (alpha helix, beta sheet)
Tertiary = 3D folding
Quaternary = spatial arrangement of protein subunits, more proteins together
Glycosylated = carbohydrate chains attached to protein backbone
1.2 History of biopharmaceuticals
Biologics (19th century) → Where it all began. Extracting medicinal products from biological sources
- Antiserum
o Tetanus antitoxin
o Diphtheria antitoxin (polyclonal antibodies from horse serum)
- Hormones/growth factors
o Insulin (1921, from pigs)
o Glucagon, FSH, Urokinase
There were problems.
- Immune reactions, contamination with infectious agents, short supply and batch-to-
batch variability
Recombinant DNA technology
Recombinant DNA technology boosted drug development in 1970s-1980s. Nowadays it is one of the
main production methods for biopharmaceuticals
Concept of recombinant DNA technology
1. mRNA strand which expresses gene of interest is isolated and cDNA is amplified using PCR
2. The cDNA sequence is transfected into expression vector
3. The expression host cells are upscaled leading to higher production of desired protein
(Upstream processing)
4. The desired proteins are recovered from the host cells and purified (Downstream
processing)
,1.3 SMD vs. BP
Small drug molecule Biopharmaceutical
+ Large diversity of chemical molecules + Any protein or nucleic acid can be used as drug
+ Well established technology + Quick discovery process
+ Cheap to manufacture + High specificity (less toxicity issues)
+ Control over chemical process is easy + Long half-life
+ Potential to cure diseases instead of alleviating
symptoms
- Discovery process is slow - Species specific
- Side effects & toxicity are common - Not orally active, don’t cross BBB, low
- Rapid elimination distribution & bioavailability
- Oral route not always an option - Longer development paths
- Potential distribution throughout the body - Expensive production
- Unstable & sensitive to heat/pH/etc.
- Control over biological process is difficult
2. Description & Registration
2.1 Classes of Biopharmaceuticals
Four major classes:
1. Extracted from living systems (animals)
a. Antibodies
b. Organs
2. Produced by recombinant DNA
a. Hormones
b. Coagulation factors & Antithrombotic factors
c. Growth factors & cytokines (interferons, interleukins)
d. (Therapeutic) monoclonal antibodies
e. Therapeutic enzymes
3. Vaccines
4. Gene therapy
a. Nucleic-acid based drugs (antisense oligonucleotides)
Hormones are used to communicate between organs and tissues for physiological regulation and
behavioural activities. They bind to specific receptor proteins in the target cell, which leads to an
activation signal transduction pathway. Commonly used hormones are oestrogens, progesterone,
and insulin.
Cytokines are a broad class of small proteins (5-20 kDa) and they play an important role in cell
signalling. They are molecules used for communication between cells to trigger the immune system
to remove pathogens. Cytokine pathways are an important target in a wide variety of diseases (auto
immune). They can be produced in vectors that are suitable for cloning (e.g. yeast cells, mammalian
cells or bacteria) when transfected with the desired gene(s), or can be modified by alteration of the
glycosylation pattern or changes in the amino acid sequence. Examples are
- Interferons
o Group of signalling proteins released in response to pathogens.
o Protect cells from virus infection
▪ Interferon beta-1a & beta-1b used to treat and control MS
▪ Can also be used to treat forms of cancer
- Interleukins
o Group of cytokines produced by white blood cells. Promote the development
and differentiation of T- and B-lymphocytes.
, ▪ Denileukin diftitox (Ontak) for cutaneous T-cell lymphoma
• Binds to an Interleukin-2 receptor and then releases diphtheria
toxin
Growth factors stimulate cell growth and proliferation in eukaryotic cells.
- Insulin-like growth factor (IGF-1)
o Polypeptide hormone similar to insulin
o Used to treat growth failure in children having trouble synthesizing IGF-1
Clotting factors are plasma glycoproteins that play a role in the blood coagulation pathway. Part of
complex chemical cascades at the site of bleeding which lead to blood clotting and finally
haemostasis.
Recombinant clotting factors are Factor VIII → Shortage leads to haemophilia A
Factor IX → shortage leads to haemophilia B
Recombinant clotting factors have an identical working mechanism to natural clotting factors:
1. The enzyme Thrombin activates recombinant Factor VIII cutting out its B-domain
2. Active recombinant Factor VIII interacts with co-factor IX and collectively activate FX
which indirectly leads to clot forming
3. Lack of either FVIII or FIX leads to excess bleeding like in haemophilia.
Enzymes catalyse chemical changes without being changed itself. They are made up of proteins.
Vaccines contain a pathogen (or a part of a pathogen) which is mostly weakened or killed. The
immune system responds and in case of next contact with the pathogen, the immune system
‘knows’ how to react and responds very quickly.
1. APCs take up and break down the antigen
2. APCs bind the peptide fragment to MHC molecule
3. APCs present the MHC antigen complex to T cells
4. T cells activate B cells
5. B cells produce antibodies
, Killed pathogens can still provoke an immune response because the immune system recognizes
PAMPs (pathogen associated molecular patterns) = molecules associated with groups of pathogens
that are recognized by cells of the innate immune system. Examples:
- Glycans, Glycoconjugates, Flagellin
- dsRNA, Bacterial lipopolysaccharides
They are recognized by Toll-Like receptors
Inactivated Vaccines
Inactivated vaccines use a killed version of the pathogen.
- Safe
- Might need several doses overtime to provide protection, because they usually don’t
provide immunity as strong as live vaccines
Live-attenuated vaccines
Live vaccines use a weakened (attenuated) version of the pathogen.
- Similar to natural infection → strong and long-lasting immune response
- 1 or 2 doses can give lifetime protection
- For people with weakened immune system, this can be dangerous
- Need to be kept cool
mRNA vaccines
Vaccines make (spike) proteins which trigger an immune response.
- Do not contain virus, so impossible to infect patient
- Short manufacturing times
Subunit, Recombinant, polysaccharide and conjugate vaccines
These vaccines use specific pieces of pathogen like its protein, sugar, or capsid.
- Give strong immune response to key parts of pathogen
- Safe for everyone (including patients with weakened immune systems)
- May need booster shots
Toxoid vaccines
Toxoid vaccines use a weakened/killed toxin made by the pathogen.
- Create immunity to the parts of a pathogen that creates the disease, not the pathogen
itself.
- Immune response is targeted to the toxin instead of the whole pathogen
- May need booster shots
Viral vector vaccines
Viral vector vaccines use a modified version of a different virus as a vector to deliver protection.
- Adenovirus is one of the most common vectors
- Delivers genetic info for the cell to make protein of the pathogen (like mRNA vaccines)
