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How Humulin Is Made?

Diabetes Is A Life-threatening Disease.

Diabetes Is A Life-threatening Disease.

Whatis Biology Good For? Controlling Diabetes: Recombinant Human Insulin (This assignment is optional and is due Friday, February 7, 2003 by noon. Read this essay and answer the questions at the bottom for 3 extra credit points. It is not necessary to visit the links in the text unless you are interested in more information.) What is Diabetes?Diabetes is a disease where the body is either unable to make the protein insulin, or is able to make insulin, but is unable to use insulin. This results in an inability to metabolize (break down) blood glucose and an inability to store glycogen (the storage form of glucose) in the liver. If left untreated, high glucose levels may damage a person's eyes, kidneys, nerves, or heart. Uncontrolled diabetes can lead to diabetic acidosis, in which toxic chemicals called ketones build up in the blood. Patients have sweet-smelling breath, and may suffer confusion, unconsciousness, and death. Diabetes is common. At least one in every 16 people has diabetes; and more than 10 million individuals with diabetes around the world rely on insulin to control their diabetes. What normally happens when a person (without diabetes) eats food? 1. Some of the food breaks down into sugars; one of these sugars is glucose, the body's main fuel. 2. Sugar enters the bloodstream, and the level of blood sugar begins to rise. 3. When the body senses an increase in blood sugar, it sends a signal to the pancreas. 4. The pancreas makes insulin and sends it into the bloodstream. 5. Insulin lowers the level of blood sugar by allowing sugar to pass from the bloodstream into the cells. 6. The level of sugar in the bloodstream falls as the sugar passes into the cells. 7. The body's cells use the sugar for fuel (ATP production). You feel energetic. When a person has diab Continue reading >>

Humulin, Manmade Insulin For Diabetics, Opening New Field Of Medical Discovery

Humulin, Manmade Insulin For Diabetics, Opening New Field Of Medical Discovery

Humulin, manmade insulin for diabetics, opening new field of medical discovery By PATRICIA McCORMACK, UPI Health Editor|Nov. 12, 1982 Some insulin-dependent diabetics are making medical history by injecting new human insulin made in the laboratory instead of the kind extracted from the pancreas glands of cattle or hogs. It is the first human healthcare product to be made by recombinant DNA technology, pioneered by Genentech Inc. of South San Francisco, Calif. The Food and Drug Administration approved the product Oct. 29. Diabetes specialists are using the new human insulin to determine the types of patients for which it is most appropriate. Diabetes is a serious, chronic disease for which there is no cure at present. But it can be controlled. It is caused by the body's failure to make use of certain foods in the normal way. Eleven-million Americans are diabetic and 10 percent have a type that is insulin-dependent. People with this kind of diabetes must take insulin by injection one or more times a day. When it is generally available and their doctors prescribe it, they will be able to use Humulin, made with DNA. DNA is short for deoxyribonucleic acid. It is DNA within each cell that carries the blueprint for life and enables species to reproduce their own kind. DNA determines whether an organism is an cat, a dog, a virus, a maple tree or a human being. Recombinant DNA technology cuts DNA molecules from two different organisms and recombines the fragments to produce biologically functional, hybrid DNA molecules. Other names for this technology are genetic engineering and gene splicing. 'The announcement of the availability in the United States of human insulin created in the laboratory with remcombinant DNA technology is an exciting event in medicine,' said Dr. Irving L Continue reading >>

Human Insulin And Recombinant Dna Technology

Human Insulin And Recombinant Dna Technology

Impaired insulin production in the beta cells of pancreas leads to the condition known as Diabetes Miletus. To treat the diabetic patient researchers produced Humulin using recombinant DNA technology by inserting human insulin gene into a vector (E. coli)... Introduction: Impaired insulin production in the beta cells of pancreas leads to the condition known as Diabetes Miletus. The bovine and porcine insulin are used to treat Patients with diabetes mellitus. But the composition of bovine and porcine insulin are slightly different from the human insulin, consequently the patient's immune system produced antibodies against animal origin insulin and also these antibodies induced inflammatory response at injection sites. Antibodies also neutralised the biological action of animal origin insulin. To overcome all these problems researchers produced Humulin using recombinant DNA technology by inserting human insulin gene into a vector (E. coli). Humulin Production Method: 1. DNA coding for A and B polypeptide chains of insulin are chemically synthesised a in the lab. Sixty three nucleotides are sequenced to produce A chain of insulin and ninety nucleotide long DNA designed to produce B chain of insulin, plus terminator codon is added at the end of each chain sequence. Anti-codon for methionine is added at the beginning of the sequence to distinguish humulin from the other bacterial proteins. 2. Chemically synthesized A and B chain DNA sequence are inserted into one of the marker gene which are present in the plasmid vector. Genes are inserted into the plasmid with the help of enzymes known as endonuclease and ligase. 3. The vector plasmids with the insulin gene are then introduced into the E. coli bacterial cell. These cells are then allowed to replicate by mitosis, along with Continue reading >>

Human Insulin

Human Insulin

Tweet Human insulin is the name which describes synthetic insulin which is laboratory grown to mimic the insulin in humans. Human insulin was developed through the 1960s and 1970s and approved for pharmaceutical use in 1982. Before human insulin was developed animal insulin, usually a purified form of porcine (pork) insulin, was used. How is human insulin produced? Human insulin is laboratory created by growing insulin proteins within E-coli bacteria (Escherichia coli). What types of human insulin are available? Human insulin is available in two forms, a short acting (regular) form and an intermediate acting (NPH) form. NPH (Neutral Protamine Hagedorn) insulin, also known as isophane insulin, is a suspension meaning that the insulin vial should be rolled or repeatedly turned upside down to ensure the solution is uniformly cloudy. Some examples of human insulin: Regular (short acting): Humulin S, Actrapid, Insuman Rapid NPH (intermediate acting): Humulin I, Insuman basal, Insulatard Premixed human insulins: Humulin M2, M3 and M5, Insuman Comb 15, 25 and 50 What are premixed human insulins? Premixed insulins consist of a mix of regular and NPH insulin. The premixed insulins are available in a number of different ratios of mixing. For example Humulin M3 is a mix of 30% short acting to 70% intermediate whereas Humulin M5 is made up of 50% of both short and intermediate acting. In recent years there has been a trend to replace human insulins with newer premixed analogue insulins. How quickly do human insulins act? Short acting (regular) insulin starts to act from about 30 minutes after injecting, with their peak action occurring between 2 and 3 hours after injecting. The duration is up to 10 hours. Intermediate acting (NPH) insulin takes about 2 to 4 hours to start acting, h Continue reading >>

Recombinant Dna Technology In The Synthesis Of Human Insulin

Recombinant Dna Technology In The Synthesis Of Human Insulin

Recombinant DNA Technology in the Synthesis of Human Insulin The nature and purpose of synthesising human insulin. Since Banting and Best discovered the hormone, insulin in 1921.(1) diabetic patients, whose elevated sugar levels (see fig. 1) are due to impaired insulin production, have been treated with insulin derived from the pancreas glands of abattoir animals. The hormone, produced and secreted by the beta cells of the pancreas' islets of Langerhans,(2) regulates the use and storage of food, particularly carbohydrates. Fig. 1 Fluctuations in diabetic person's blood glucose levels, compared with healthy individuals. Source: Hillson,R. - Diabetes: A beyond basics guide, pg.16. Although bovine and porcine insulin are similar to human insulin, their composition is slightly different. Consequently, a number of patients' immune systems produce antibodies against it, neutralising its actions and resulting in inflammatory responses at injection sites. Added to these adverse effects of bovine and porcine insulin, were fears of long term complications ensuing from the regular injection of a foreign substance,(3) as well as a projected decline in the production of animal derived insulin.(4) These factors led researchers to consider synthesising Humulin by inserting the insulin gene into a suitable vector, the E. coli bacterial cell, to produce an insulin that is chemically identical to its naturally produced counterpart. This has been achieved using Recombinant DNA technology. This method (see fig. 2) is a more reliable and sustainable(5) method than extracting and purifying the abattoir by-product. Fig. 2 An overview of the recombination process. Source: Novo - Nordisk promotional brochure,pg 6. Understanding the genetics involved. The structure of insulin. Chemically, insuli Continue reading >>

How Did They Make Insulin From Recombinant Dna?

How Did They Make Insulin From Recombinant Dna?

Recombinant DNA is a technology scientists developed that made it possible to insert a human gene into the genetic material of a common bacterium. This “recombinant” micro-organism could now produce the protein encoded by the human gene. Continue reading >>

A New Insulin Given Approval For Use In U.s.

A New Insulin Given Approval For Use In U.s.

Archives |A NEW INSULIN GIVEN APPROVAL FOR USE IN U.S. , Page 001001 The New York Times Archives The Food and Drug Administration yesterday approved the marketing of human insulin made artificially from gene-splicing techniques. It is the first such product of genetic engineering to be granted Governmental approval for human use. The new insulin, called Humulin, is manufactured by a technique known as recombinant DNA, which involves inserting human genetic instructions into a bacterium that then produces the drug. The approval by the Food and Drug Administration came five months after application was made by Eli Lilly and Company of Indianapolis, Ind. Normally, such approval takes 20 to 30 months. The efforts to produce the artificial human hormone commercially began four years ago, after scientists at the City of Hope National Medical Center in Duarte, Calif., and Genentech Inc. in South San Francisco first succeeded in producing it in the laboratory. The new insulin is expected by its manufacturer to be available in drug stores without a prescription sometime in 1983. A spokesman for Lilly said Humulin was expected to cost about twice as much initially as the animal insulins now used by an estimated two million of the 10 million diabetics in the United States. Called a Major Step Forward Dr. Henry Miller, the medical officer in charge of Humulin at the F.D.A., said the development was a major step forward in the ''scientific and commercial viability of'' recombinant DNA techniques. ''We have now come of age,'' Dr. Miller said. Dr. Irving L. Spratt, president of the American Diabetes Association, said: ''The announcement is an exciting event in medicine. It demonstrates the melding of intensive research in genetic engineering with complex pharmaceutical production.'' Continue reading >>

How Insulin Is Made - Material, Manufacture, History, Used, Parts, Components, Structure, Steps, Product

How Insulin Is Made - Material, Manufacture, History, Used, Parts, Components, Structure, Steps, Product

Background Insulin is a hormone that regulates the amount of glucose (sugar) in the blood and is required for the body to function normally. Insulin is produced by cells in the pancreas, called the islets of Langerhans. These cells continuously release a small amount of insulin into the body, but they release surges of the hormone in response to a rise in the blood glucose level. Certain cells in the body change the food ingested into energy, or blood glucose, that cells can use. Every time a person eats, the blood glucose rises. Raised blood glucose triggers the cells in the islets of Langerhans to release the necessary amount of insulin. Insulin allows the blood glucose to be transported from the blood into the cells. Cells have an outer wall, called a membrane, that controls what enters and exits the cell. Researchers do not yet know exactly how insulin works, but they do know insulin binds to receptors on the cell's membrane. This activates a set of transport molecules so that glucose and proteins can enter the cell. The cells can then use the glucose as energy to carry out its functions. Once transported into the cell, the blood glucose level is returned to normal within hours. Without insulin, the blood glucose builds up in the blood and the cells are starved of their energy source. Some of the symptoms that may occur include fatigue, constant infections, blurred eye sight, numbness, tingling in the hands or legs, increased thirst, and slowed healing of bruises or cuts. The cells will begin to use fat, the energy source stored for emergencies. When this happens for too long a time the body produces ketones, chemicals produced by the liver. Ketones can poison and kill cells if they build up in the body over an extended period of time. This can lead to serious illne Continue reading >>

Recombinant Dna Used To Produce Human Insulin

Recombinant Dna Used To Produce Human Insulin

Humulin Production: Ever since Banting and Best isolated a protein they called "isletin" from the Islets of Langerhans from a pancreas and injected it into diabetic dogs, insulin has been used to help diabetics live out their lives. However, this protein had its problems. For many years, insulin was extracted and purified from either porcine or bovine pancreases, and this carried with it two main difficulties. The first was that though the animal insulin was chemically similar to human insulin, there were some differences, and these differences led to antibody attack and inactivation as well as inflammation in many patients. Also, there was the problem that this method of extracting insulin from animal organs made it difficult to obtain large amounts of pure insulin. Nowadays, it is a well known fact that DNA contains genes which encode all the different proteins that an organism can produce. But back then, the composition of proteins and their relation to DNA was unknown. It wasn't until Sanger, in 1955, determined the sequence of insulin and determined that proteins are composed of specific amino acids linked to each other in a peptide chain. And even then, it wasn't until 1979 that human insulin was able to be produced in large quantities. This was done by the use of recombinant DNA. Recombinant DNA, or rDNA, is DNA which specifically encodes a protein. This is cut from genomic DNA by a restriction enzyme which cuts DNA at specific sequences along the chain. These pieces are then analyzed and the DNA needed to make the protein is extracted and purified. Since insulin contains two polypeptide chains linked by disulfide bonds, two pieces of DNA are extracted. These DNA strands are then placed into two different plasmids, as shown in the figure below. These plasmids are Continue reading >>

Humulin N

Humulin N

HUMULIN® N (human insulin [rDNA origin]) isophane) Suspension DESCRIPTION HUMULIN N (human insulin [rDNA origin] isophane) suspension is a human insulin suspension. Human insulin is produced by recombinant DNA technology utilizing a non-pathogenic laboratory strain of Escherichia coli. HUMULIN N is a suspension of crystals produced from combining human insulin and protamine sulfate under appropriate conditions for crystal formation. The amino acid sequence of HUMULIN N is identical to human insulin and has the empirical formula C257H383N65O77S6 with a molecular weight of 5808. HUMULIN N is a sterile white suspension. Each milliliter of HUMULIN N contains 100 units of insulin human, 0.35 mg of protamine sulfate, 16 mg of glycerin, 3.78 mg of dibasic sodium phosphate, 1.6 mg of metacresol, 0.65 mg of phenol, zinc oxide content adjusted to provide 0.025 mg zinc ion, and Water for Injection. The pH is 7.0 to 7.5. Sodium hydroxide and/or hydrochloric acid may be added during manufacture to adjust the pH. Continue reading >>

Dave Groves [mailto:dggroves@earthlink.net]

Dave Groves [mailto:[email protected]]

From: Dave Groves [mailto:[email protected]] Sent: Monday, February 18, 2002 2:15 PM Subject: Insulin Information – How is “Human” insulin made? There are many ways to produce "human" insulin and I will outline my understanding of the various means and who is reported to be using which method(s). 1. Natural human insulin can be extracted, crystallized, and purified from cadaverous human pancreas tissue as it is from beef pancreas and pig pancreas. No one does this, as the few human pancreases available are far to needed for pancreas transplantation, islet cell transplantation, and other research for curing diabetes. Such insulin would likely cost well over $5,000 per vial (it would require well over 3 cadavers per vial) and there would be almost no supply, perhaps enough for 10 diabetics in the world. It would be no better than any other "human" insulin and it would be subject to most, if not all, of the problems that we know exist for many of us with the semi-synthetic, synthetic, and natural animal insulins. 2. Semi-synthetic human insulin can be made, biochemically, from natural pork insulin, by removing the B30 amino acid (Alanine) from pork insulin and substituting a Theronine amino acid in its place. For all vertebrate animals, each insulin molecule consists of precisely 2 peptide chains (A and B) bound together by sulfa bonds at the A7-B7 Cysteine site and at the A20-B19 Cysteine site and there is an additional Cysteine sulfa bond at the A6-A11. All insulin molecules consist of this two chain structure, with an A chain of 21 amino acids and a B chain of 30 amino acids, for a total of 51 amino acid molecules bound by 3 sulfa bonds, any molecule with more or fewer amino acids or different locations for the 3 sulfa bonds cannot be insulin, though it may b Continue reading >>

Regular Insulin

Regular Insulin

Regular insulin, also known as neutral insulin and soluble insulin is a type of short acting insulin.[1] It is used to treat diabetes mellitus type 1, diabetes mellitus type 2, gestational diabetes, and complications of diabetes such as diabetic ketoacidosis and hyperosmolar hyperglycemic states.[3] It is also used along with glucose to treat high blood potassium levels.[4] Typically it is given by injection under the skin, but may also be used by injection into a vein or muscle.[1] Onset of effect is typically in 30 minutes and they last for 8 hours.[3] The common side effect is low blood sugar.[3] Other side effects may include pain or skin changes at the sites of injection, low blood potassium, and allergic reactions.[3] Use during pregnancy is relatively safe for the baby.[3] Regular insulin can be made from the pancreas of pigs or cows.[1] Human versions can be made either by modifying pig versions or recombinant technology.[1] Insulin was first used as a medication in Canada by Charles Best and Frederick Banting in 1922.[5] It is on the World Health Organization's List of Essential Medicines, the most effective and safe medicines needed in a health system.[6] The wholesale cost in the developing world is about 2.39 to 10.61 USD per 1,000 iu of regular insulin.[7] In the United Kingdom 1,000 iu costs the NHS 7.48 pounds, while in the United States this amount is about 134.00 USD.[1][8] Versions are also available mixed with longer–acting versions of insulin, such as NPH insulin.[1] Medical uses[edit] It is used for the long term management of diabetes.[3] Regular insulin is the treatment of choice for the two diabetic emergencies diabetic ketoacidosis and hyperosmolar hyperglycemic states.[3] It may also be used in combination with glucose to lower potassium leve Continue reading >>

First Successful Laboratory Production Of Human Insulin Announced

First Successful Laboratory Production Of Human Insulin Announced

South San Francisco, Calif. -- September 6, 1978 -- Genentech, Inc. and City of Hope National Medical Center, a private research institution and hospital in Duarte, California today announced the successful laboratory production of human insulin using recombinant DNA technology. Insulin is a protein hormone produced in the pancreas and used in the metabolism of sugar and other carbohydrates. The synthesis of human insulin was done using a process similar to the fermentation process used to make antibiotics. The achievement may be the most significant advance in the treatment of diabetes since the development of animal insulin for human use in the 1920's. The insulin synthesis is the first laboratory production DNA technology. Recombinant DNA is the technique of combining the genes of different organisms to form a hybrid molecule. DNA (deoxyribonucleic acid), the substances genes are composed of, contains the chemical record in which genetic information is encoded. Scientists at Genentech and City of Hope inserted synthetic genes carrying the genetic code for human insulin, along with the necessary control mechanism, into an E. coli bacterial strain which is a laboratory derivative of a common bacteria found in the human intestine. Once inside the bacteria, the genes were "switched-on" by the bacteria to translate the code into either "A" or "B" protein chains found in insulin. The separate chains were then joined to construct complete insulin molecules. The development of genetically engineered human insulin was funded by Genentech. However, the work was a cooperative effort between Genentech and City of Hope. The synthesis of human insulin gene was accomplished by four scientists at City of Hope Medical Center led by Roberto Crea, Ph.D., and Keichi Itakura, Ph.D. Scien Continue reading >>

Humulin

Humulin

About Humulin Humulin is human insulin, unlike the insulin you have used in the past, Humulin does not come from the pancreas of any animal. And though it is, in fact, human insulin, Humulin does not come from the human pancreas -- which produces insulin in people who do not have diabetes. You are probably wondering, "Then where does it come from?" Humulin is made by a special process called recombinant DNA (rDNA) technology. This process makes it possible to produce insulin identical to your body's own. And, more than likely, this is the reason your doctor is giving you Humulin. Will you need to change your dosage? Most patients transfer to Humulin at the same dose and dosage schedule used with their previous insulin. Your doctor may have to make some dosage adjustments over a period of weeks or months, however, depending on how you respond to Humulin. What other changes must you make? As you know, diet and exercise are very important for good control of your blood sugar. Your doctor or nurse-educator may recommend some changes in your diet or exercise program if your control has been less than satisfactory. The fact that you are on Humulin does not make diet and exercise any less important. How do you use Humulin? Follow the instructions of your doctor or diabetes nurse educator.* Read "Information for the Patient" that accompanies your vial of Humulin Remember to test your blood glucose as directed by your doctor or nurse educator. If you have any questions or are not feeling well, call your doctor or nurse educator. Any change of insulin should be made cautiously and only under medical supervision. Changes in refinement, purity, strength, brand (manufacturer), type (regular, NPH, Lente, etc), species (beef, pork, beef-pork, human), and/or method of manufacture (reco Continue reading >>

Humulin : Synthetic Insulin

Humulin : Synthetic Insulin

Humulin is synthetic human insulin prepared by using genetic engineering. Humulin is manufactured from DNA sources in laboratory, using recombinant DNA technology. Synthetic insulin is also called genetically engineered insulin. The synthetic insulin (humulin) is as effective as hormone insulin secreted by human pancreas. What is Humulin? Humulin is synthetic human insulin prepared by using genetic engineering. Humulin is manufactured from DNA sources in laboratory, using recombinant DNA technology. Synthetic insulin is also called genetically engineered insulin. The synthetic insulin (humulin) is as effective as hormone insulin secreted by human pancreas. Synthesis of Humulin In 1978, scientists synthesized human insulin from E.coli bacteria using recombinant DNA technology, by preparing two DNA sequences for A and B chains of human insulin and introduced them in plasmid of E.coli. This led to production of human insulin chain. Eli Lilly, an American company marketed the first human insulin called humulin in 1983. Eli Lilly and Ranbaxy launched a new insulin project namely Humalog (an analog of 5, 6 human insulin), which is more expensive than human insulin products, but have good absorption in body, as compared to other insulin products. Genetically engineered Insulin : Humulin, preferred over old animal based products Structure of Insulin Insulin is a proteinaceous hormone secreted by beta-cells of islets of langerhans of pancreas. Insulin controls, blood sugar level and when there is less secretion of insulin, it results in diabetes (high blood- sugar level). In 1954, Frederick Sanger determined primary structure of Insulin. Insulin is a protein formed by two polypeptide chains: A-chain and B-chain, interlinked by two sulphide bonds (see fig). A-chain is formed of 2 Continue reading >>

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