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Lecture 30

MBIO 1010 Lecture 30: Lecture 30

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University of Manitoba
MBIO 1010
Christopher Rathgeber

Microbiology 1010 – Lecture Notes Manufacturing Human Insulin Through research, Frederick Banting discovered mechanism of diabetes   Insulin was 1 produced in 1921 by F. Banting and Charles Best harvested insulin from animal (dog) pancreas  1923 Eli Lilly began large-scale commercial production  Ground up bovine and swine pancreas glands Animal Insulin has limitations  Although similar to human insulin, bovine and porcine insulin are not the same Many patients form antibodies against the foreign protein   Causes inflammation and allergic responses  Fear of long-term affects of using foreign protein Stimulated search for new ways of making HUMAN insulin in large volumes  Solution: insert human insulin gene into E. coli  produce human insulin – recombinant DNA technology Body sees it as foreign when it comes into the body – so we destroy it with an immune response. Structure of Insulin  Insulin – protein hormone 51 amino acids  Two chains: o α chain – 21 a.a. β chain – 30 a.a. Held together by disulfide bridges Bacteria still have limitations … simple polypeptide is ok... But anything else too complex is not. Manufacturing Human Insulin Synthesis of DNA containing the nucleotide sequences of the A and B polypeptide chains of insulin 158 Microbiology 1010 – Lecture Notes Manufacturing Human Insulin (Continued…) DNA containing the A-chain gene and B-chain gene individually cloned into a plasmid vector   The recombinant plasmids DNA are then introduced into E. coli  Bacteria containing the plasmid grown as large-scale cultures in fermenters become insulin factories that produce large amounts of A-chain and B-chain polypeptide Introduce the cloned gene into E.coli Manufacturing Human Insulin (Continued…) Extraction and purification of A and B chains   A- and B-chains connected via disulfide bridges Results in pure synthetic human insulin Have the insulin but now you have to get rid of the contaminates Human because it was made from a human recipe Synthetic = because it was not made within the body 159 Microbiology 1010 – Lecture Notes Radioactive bacteria reduce the spread of pancreatic cancer Pancreatic Cancer o Highly aggressive o Poor survival rate - <4% of people survive 5 years after diagnosis o “Silent killer” - cancer spreads (metastasizes) before primary tumor detected o Patients with metastasized cancer – jaundice, pain, weight loss, fatigue o At this point incurable – treatment to improve quality of life only o Chemotherapeutic agents currently in use – improve median life expectancy 6 months o Urgent need for alternative therapies Extremely vigorous= because it is not detected until it is too late. You can’t eliminate – survival is less than 10% Patients that have this cancer is usual symptoms of weight loss (not same digestion – indigestion of food) Causes pain and fatigue as well. Chemo – only increases life for 6 months of less What has been shown to be effective is a bug Listeria monocytogenes – a promising alternative  Weakened lab form (= attenuated) of L.m. can infect cancer cells ≠ healthy cells Normal t
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