Quiz 5 Flashcards
(99 cards)
biotechnology
technique which uses living organisms (microorganisms) bacteria, yeast, mammalian in the production of products used to affect human health and human environment
uses of biotech
treatment, prevention, diagnostics
size of biotech
macromolecules
biotech products
proteins, Nucleic acids, monoclonal antibodies, RNA
small molecule medicines vs large molecule medicines
small: single chem synthesized, active ingredient, made entirely from chem-synthesized reactions between different compounds, manufactured in a chemical process
large: made from living cells and complex, active ingredients are protiens, antibodies, cytokines, insulin, biologics derived from living organisms, characteristics and properties influenced by the manufacturing process, sensitive to changes in their enviroment and handling (all different final products)
CDER vs CBER
both FDA regulated CDER is for small molecule medicines and CBER is for large molecule medicines
first biotech product
insluin, 1921, university of toronto, banting/best
insulin then vs now
then: isolated from cows and pigs
now: recombinant human insulin
issues with then: animals not all the same, allergies/immune response, containmenents
rDNA
used to produce biologics, proteins, mAbs, developed in 1973, used to obtain large amounts of protein higher level of purity and lower cost
PCR
polymerase chain reaction, proteins, gene therapy, antisense NAs, large scale production if possible
hybridoma technology
antibody production
when was the first rDNA marketed?
insulin - 1982 - FDA approved
how to obtain the biotech product / protien
- isolation of DNA with gene of interest
- insertion into plasmid for protein synthesis - rDNA - independent of nucleus
- host selection for scale-up
cohen-boyer method
1971-EcoRI sed to cut plasmid restriction endonuclease
1972 -insertion of rDNA so foreign DNA will replicate naturally
step 1 cohen boyer method
DNA isolation - DNA first cut into smaller lengths with restriction endonucleases which recognize specific sequences of base pairs and cut the DNA at that point. the DNA sequence desired can therefore be removed and isolated.
step 2 cohen boyer method
recombinant DNA production - protein production begins when incorporating the DNA of interest into the plasmid DNA, DNA segment mixed with the plasmid DNA and ligase, ligase connects the DNA with plasmid, forms the rDNA
step 3 cohen boyer method
host cell selection and protein production - cloning accomplished by inserting the recombinant DNA into a host that replicates easily - bacteria, yeast, mammalian cells, proteins increase in complexity with increasing host complexity
bacterial hosts
advantages: replication rate is rapid, cheap used for simple proteins
disadvantages: bacterial debris, pyrogens, antigens, fever causing not in here!
cannot make relevant post-translational modifications
can do glycosylation
recombinant insulin came from
E. Coli
humalin
rDNA insulin
yeast host cells
advantages: protien secretion, growth rate, large-scale production, absence, not pathogenic, post-translational modifications
disadvantages: active proteases can degrade proteins
yeasts are attractive hosts for the production of therapuetic proteins, used to express recombinant proteins to overcome the shortcomings of bacterial expression systems
example of yeast host
saccaromyces cerevisiae, leukine is a drug
mammalian host cells
some proteins only produced with higher organisms, proteins are difficult to express and need folding complex for function
advantages: folding, post-translational modifications, contamination, more complex proteins
disadvantages: higher cost, more time
example of mammalian host cells
chinese hamster ovary cells, aransep is an EPO produced in CHO
