Mid Term review.docx - Mid Term review Lecture 1 Biopharmaceuticals Medicines from recombinant DNA technology You have gene and DNA Once you know gene

Mid Term review.docx - Mid Term review Lecture 1...

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Mid Term review Lecture 1: Biopharmaceuticals: Medicines from recombinant DNA technology - You have gene and DNA - Once you know gene or DNA then you can make the gene to be useful into a protein - - first you have to insert gene into plasmid - Put DNA of interest into plasmid first then plasmid into host cell line – transfection - Bioreactor – - Take it from super natant - Cell purification and collect product in supernatant then purification - After you purify you formulate and fill Since 1980s many biopharmaceuticals got approval as medicines - First product that came out from recombinant was insulin or Humulin R - Second product was protropin - Insulin was purified from animals o It’s a simple protein and this was the beginning of recombinant proteins - Later on came the human growth hormone and - Most drugs were enzymes and proteins, antibodies were developed later on Host cell options and microbial systems Genetic Engineered organisms to produce biopharma: host options: Bacteria cell, Animal cell, Plant cell First generation of biopharma used microbial host (e.coli) - Bacterial used for protein production o E.coli: major host o High protein expression o Rapid growth rate: high productivity o Ease of manufacturing o Low cost: Medium for growth and product Disadvantages of e. coli - Intra cellular expression: not secreted - protein aggregated: biologically inactive - Must resort to in vitro protein refolding Yeast is more complicated than e coli - Yeast can produce insulin Microbial Hosts: E. Coli., Saccharomyces cerevisiae, Pichia pastoris
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Cell culture technology Cell Culture Technology: More complex biopharmaceuticals require sophisticated expression systems and well-controlled processes - Biological activity requires : correct folding, correct bridging, modifications to the amino acid residues, glycosylation, sialyation, etc - Most of these properties cant be predicted by the protein sequence - Manufacturing process conditions influence the structure of the molecule Manufacturing process product quality Post translational modifications of proteins: glycosylation and sialyation - Folding sugar bonds - Cells assemble proteins - Manufacturing process makes the product Sialyation of the proteins: role of terminal sialic acid on half life - You can control this by the environment of the cells - Presence of sialic acid affects circulatory half life time for therapeutic glycoproteins Cell culture is needed for complex biopharmaceuticals - Advantages of cell culture systems o Product secreted: active biologically o Capable of post-translational modification Are everything that goes into the protein after the protein is assembled in the peptide o Glycosylation provides Higher biological activity Greater solubility Longer half-life via sialyation - Disadvantages of cell culture systems o Lower growth rate: lower productivity o Complex nutrition and higher raw material cost o More difficult in manufacturing Cell strains and lines for biopharma production - Primary cells: cells coming from tissue directly o
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