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Application of biotech Xingzhi Wang School of Life Sciences Northeast Normal University.

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1 Application of biotech Xingzhi Wang School of Life Sciences Northeast Normal University

2 History of Biotechnology The term "biotechnology" was coined in 1919 by Karl Ereky, an Hungarian engineerbiotechnology Traditional biotechnology has been used for thousands of years to produce improved food and health care products. Today, modern biotechnology enables us to develop improved products more safely and more rapidly than ever before.

3 Examples Examples:  they could plant their own crops and breed their own animals, they learned to use biotechnology.  The discovery that fruit juices fermented into wine, or that milk could be converted into cheese or yogurt, or that beer could be made by fermenting solutions of malt and hops began the study of biotechnology

4 Examples cont ’ d  When the first bakers found that they could make a soft, spongy bread rather than a firm, thin cracker, they were acting as fledgling biotechnologists.  The first animal breeders, realizing that different physical traits could be either magnified or lost by mating appropriate pairs of animals, engaged in the manipulations of biotechnology.

5 Definition Definition: What then is biotechnology? the term "biotechnology" refers to the use of living organisms or their products to modify human health and the human environment. Other definitions: 1. Biotechnologists use engineering and science to create new products from biologically based raw materials, such as vaccines or foods. They also develop factory processes to reduce pollution or treat waste products. 2. Biotechnology uses living cells and materials produced by cells to create pharmaceutical, diagnostic, agricultural, environmental, and other products to benefit society.

6 Introduction Biotechnology is essentially  the use of living organisms (often minute microorganisms) and their products  for health, social or economic purposes. Biotechnology is widely considered to be the growth technology of the 21st Century which will lead to huge growth in the Biotechnology industry and exciting opportunities for graduates.

7 BIOTECNOLOGY FIELDS OF APPLICATION

8 Introduction Through genetic engineering scientists can combine DNA from different sources and this process is called “ Recombinant DNA technology ” The secrets of DNA structure and functions have led to gene cloning and genetic engineering, manipulating the DNA of an organism

9 Introduction Biotechnology is an interdisciplinary science including not only biology, but also subjects like mathematics, physics, chemistry, computing and engineering. It is a blend of various technologies applied together to living cells for production of a particular product or for improving upon it.

10 BIOTECNOLOGY BIOCHEMISTRY MICROBIOLOGY GENETICS PHYSIOLOGY EMBRIOLOGY INMUNOLOGY MOLECULAR BIOLOGY CELL BIOLOGY ANALYTICAL CHEMISTRY CHEMICAL ENGINEERING INFORMATICS

11 Nature of Work The nature of work of biotechnologists, being interdisciplinary, requires working together of people from different fields such as biology, chemistry, biochemistry, microbiology, molecular biology, immunology, genetics, engineering, food science, agriculture etc.

12 Periods of Biotechnology History Pre- 1800: Early applications and speculation : Significant advances in basic understanding : Genetics : DNA research, science explodes present: modern biotechnology

13 Biotechnology Time Lines 6000 BC Yeast was used to make beer by Sumerians and Babylonians BC The Egyptians discovered how to bake leavened bread using yeast. 420 BC Socrates (470? BC), the Greek philosopher, speculated on why children don't always resemble their parents. 320 BC Aristotle ( BC), told his students that all inheritance comes from the father.

14 Biotechnology Time Lines 1000 AD Hindus observed that certain diseases may "run in the family." Spontaneous Generation is the dominant explanation that organisms arise from non-living matter. Maggots, for example, were supposed to arise from horsehair.Spontaneous Generation 1630 AD William Harvey concluded that plants and animals alike reproduce in a sexual manner: – egg isolated in 1800 ’ sWilliam Harvey AD Marcello Malpighi ( ) in this period used a microscope to study blood circulation in capillaries, described the nervous system as bundles of fibers connected to the brain by the spinal cord.

15 Biotechnology Time Lines 1673 AD Anton van Leeuwenhoek ( ), He was the first scientist to describe protozoa and bacteria and to recognize that such microorganisms might play a role in fermentation Giacomo Pylarini in Constantiople practiced "inoculation"--intentionally giving children smallpox to prevent a serious case later in life. Inoculation will compete with "vaccination"--an alternative method that uses cowpox rather than smallpox as the protecting treatment--for a century.

16 Biotechnology Time Lines 1809 Nicolas Appert devised a technique using heat to can and sterilize food 1827 The worldwide search for the elusive mammalian egg ended with the first observation of canine eggs. Remember 1630 and William Harvey 1850 ONE OF MY FAVORITES Ignaz Semmelweis used epidemiological observations to propose the hypothesis that childbed fever can be spread from mother to mother by physicians. He tested the hypothesis by having physicians wash their hands after examining each patient. He became despised by the medical profession and lost his job.

17 Biotechnology Time Lines 1856 Karl Ludwig discovered a technique for keeping animal organs alive outside the body, by pumping blood through them. Louis Pasteur ( ) asserted that microbes are responsible for fermentation. Louis Pasteur 1859 Charles Darwin ( ) hypothesized that animal populations adapt their forms over time to best exploit the environment, a process he referred to as "natural selection." As he traveled in the Galapagos Islands, he observed how the finch's beaks on each island were adapted to their food sources.

18 Biotechnology Time Lines 1863 Louis Pasteur invented the process of pasteurization, heating wine sufficiently to inactivate microbes (that would otherwise turn the "vin" to "vin aigre" or "sour wine") while at the same time not ruining the flavor of the wine ℃ for half hour.Louis Pasteur Anton de Bary proved that a fungus causes potato blight. A challenge for scientists during this period was to discern whether a microbe was the cause of, or the result of, a disease.

19 Biotechnology Time Lines 1865 Gregor Mendel ( ), an Augustinian monk, presented his laws of heredity to the Natural Science Society in Brunn, Austria. Mendel proposed that invisible internal units of information account for observable traits, and that these "factors" - which later became known as genes - are passed from one generation to the next. Mendel's work remained unnoticed, languishing in the shadow of Darwin's more sensational publication from five years earlier, until 1900, when Hugo de Vries, Erich Von Tschermak, and Carl Correns published research corroborating Mendel's mechanism of heredity..Gregor Mendel

20 Biotechnology Time Lines Pasteur investigated silkworm disease and established that diseases can be transmitted from one animal to another. Pasteur 1865 Joseph Lister began using disinfectants such as phenol (=carbolic acid) in wound care and surgery as Pasteur developed the germ theory of disease Pasteur

21 Biotechnology Time Lines 1868 Davaine used heat treatment to cure a plant of bacterial infection. Fredrich Miescher, a Swiss biologist, successfully isolated nuclein, a compound that includes nucleic acid, from pus cells obtained from discarded bandages W. Flemming discovered mitosis.

22 Biotechnology Time Lines 1871 DNA was isolated from the sperm of trout found in the Rhine River Robert Koch investigated anthrax and developed techniques to view, grow, and stain organisms. He then photographed them, aided by Gram, Cohn, and Weigart Studying fowl cholera, Pasteur published his work on "attenuated" or weakened strains of organisms that could not cause disease but protected against severe forms of the same disease.Pasteur

23 Biotechnology Time Lines 1881 ANTRAX Robert Koch described bacterial colonies growing on potato slices, on gelatin medium, and on agar medium. Nutrient agar became a standard tool for obtaining pure cultures and for identifying genetic mutants. This is considered by T.D. Brock to be the single most important discovery in the rise of microbiology. Pasteur used attenuation to develop vaccines against the bacterial pathogens of fowl cholera and anthrax; this was a founding moment in immunology and opened new areas in the field of preventive medicine.

24 Biotechnology Time Lines 1883 Emil Christian Hansen made the first pure yeast culture for beer production in Gamle Carlsberg Brewery, Copenhagen, Denmark.

25 Biotechnology Time Lines 1884 ROBERT KOCH STATED HIS "POSTULATES" FOR TESTING WHETHER A MICROBE IS THE CAUSAL AGENT OF A DISEASE. Pasteur developed a rabies vaccine. Pasteur Christian Gram described the differential staining technique for bacteria known as the Gram stain. Gregor Mendel died after 41 years of meticulously studying the heredity "factors" of pea plants. Having received no scientific acclaim during his lifetime, he said not long before his death, "My time will come." Gregor Mendel

26 Biotechnology Time Lines Converging on DNA 1900 MENDEL ’ S WORK FINALLY TOOK ON IMPORTANCE The science of genetics was finally born when Mendel's work was rediscovered by three scientists - Hugo DeVries, Erich Von Tschermak, and Carl Correns - each one independently researching scientific literature for precedents to their own "original" work.Mendel's 1902 HUMAN GENETICS BORN Walter Stanborough Sutton stated that chromosomes are paired and may be the carriers of heredity. He suggested that Mendel's "factors" are located on chromosomes.

27 Biotechnology Time Lines 1905 X AND Y CHROMOSOMES RELATED TO GENDER Edmund Wilson and Nellie Stevens proposed the idea that separate X and Y chromosomes determine sex. They showed that a single Y chromosome determines maleness, and two copies of the X chromosome determine femaleness William Bateson and Reginald Crudell Punnett, along with others, demonstrated that some genes modify the action of other genes Paul Erlich investigated atoxyl compounds and discovered the beneficial properties of Salvarsan - the first chemotherapeutic agent.

28 Biotechnology Time Lines 1907 Thomas Hunt Morgan began his work with fruit flies that will prove that chromosomes have a definite function in heredity, establish mutation theory, and lead to a fundamental understanding of the mechanisms of heredity MENDEL ’ S LAWS TO ANIMALS Wilhelm Johannsen coined the terms 'gene' to describe the carrier of heredity; 'genotype' to describe the genetic constitution of an organism; and 'phenotype' to describe the actual organism, which results from a combination of the genotype and the various environmental factors.

29 Biotechnology Time Lines 1910 BASIS OF MODERN GENETICS Thomas Hunt Morgan proved that genes are carried on chromosomes, establishing the basis of modern genetics. With his co-workers, he pinpointed the location of various fruit fly genes on chromosomes, establishing the use of Drosophila fruit flies to study heredity Thomas Hunt Morgan explained the separation of certain inherited characteristics that are usually linked as caused by the breaking of chromosomes sometimes during the process of cell division. Morgan began to map the positions of genes on chromosomes of the fruit fly.

30 Biotechnology Time Lines 1912 Lawrence Bragg discovered that X-rays can be used to study the molecular structure of simple crystalline substances Herbert M. Evans found (incorrectly) that human cells contain 48 chromosomes EUGENICS IN THE UNITED STATES Politicians encouraged by the eugenics movement passed the U.S. Immigration Act of 1924, limiting the influx of poorly educated immigrants from Southern and Eastern Europe on the grounds of suspected genetic inferiority.

31 Biotechnology Time Lines 1926 Thomas Hunt Morgan published 'The theory of the gene', the culmination of work on the physical basis for Mendelian genetics based on breeding studies and optical microscopy. Hermann Muller discovered that X-rays induce genetic mutations in fruit flies 1,500 times more quickly than under normal circumstances. This discovery provided researchers with a way to induce mutations, an important tool for discovering what genes do on their own.

32 Biotechnology Time Lines 1928 Fredrick Griffiths noticed that a rough type of bacterium changed to a smooth type when an unknown "transforming principle" from the smooth type was present. Sixteen years later, Oswald Avery identified that "transforming principle" was DNA. Alexander Fleming noticed that all the bacteria in a radius surrounding a bit of mold in a petri dish had died. The age of penicillin thus began, although it would be almost 15 years before it was made available to the community for medicinal use.

33 Biotechnology Time Lines 1938 Proteins and DNA were studied in various labs with X-ray crystallography. The term "molecular biology" was coined ONE GENE ONE ENZYME George Beadle and Edward Tatum experimented with Neurospora, a mold that grows on bread in the tropics, developing the "one-gene-one- enzyme" hypothesis: each gene is translated into an enzyme to perform tasks within an organism.

34 Biotechnology Time Lines 1943 The Rockefeller Foundation, collaborating with the Mexican government, initiated the Mexican Agricultural Program. This was the first use of plant breeding as foreign aid Cortisone was first manufactured in large amounts. KIND OF A FIRST BIOTECH PRODUCT 1944 Waksman isolated streptomycin, an effective antibiotic for TB.

35 Biotechnology Time Lines 1945 The U.N. Food and Agriculture Organization (FAO) was formed in Quebec, Canada CELLS GROWN IN LAB Isolated animal cell cultures were grown in laboratories Barbara McClintock first reported on "transposable elements" - known today as "jumping genes." The scientific community failed to appreciate the significance of her discovery at the time. Barbara McClintock

36 Biotechnology Time Lines 1950 Erwin Chargaff found that in DNA the amounts of adenine and thymine are about the same, as are the amounts of guanine and cytosine. These relationships are later known as "Chargaff's Rules" and serve as a key principle for Watson and Crick in assessing various models for the structure of DNA. AT ABOUT THE SAME; GC ABOUT THE SAME.

37 Biotechnology Time Lines : Expanding the Boundaries of DNA Research The discovery of the structure of DNA resulted in an explosion of research in molecular biology and genetics, paving the way for the biotechnology revolution Nature magazine published James Watson's and Francis Crick's manuscript describing the double helix structure of DNA.James Watson's

38 Biotechnology Time Lines 1953 Gey developed the HeLa human cell line. HENRIETTA LACKS- DIED IN 1951 OF CERVICAL CANCER- MOTHER OF 5- HER CELLS FIRST SHOWN TO GROW OUTSIDE THE BODY FOR EXTENDED PERIODS- USED TO DEVELOP THE POLIO VACCINE

39 Biotechnology Time Lines 1957 CENTRAL DOGMA OF DNA- HOW DNA MAKES A PROTEIN Francis Crick and George Gamov worked out the "central dogma," explaining how DNA functions to make protein Francois Jacob and Jacques Monod established the existence of genetic regulation - mappable control functions located on the chromosome in the DNA sequence - which they named the repressor and operon.

40 Biotechnology Time Lines 1962 Watson and Crick shared the 1962 Nobel Prize for Physiology and Medicine with Maurice Wilkins. Unfortunately, Rosalind Franklin, whose work greatly contributed to the discovery of the double helical structure of DNA, died before this date, and the Nobel Prize rules do not allow a prize to be awarded posthumously. Rosalind Franklin

41 Biotechnology Time Lines 1966 GENETIC CODE CRACKED The genetic code was "cracked". Marshall Nirenberg, Heinrich Mathaei, and Severo Ochoa demonstrated that a sequence of three nucleotide bases (a codon) determines each of 20 amino acids.

42 Biotechnology Time Lines 1967 Arthur Kornberg conducted a study using one strand of natural viral DNA to assemble 5,300 nucleotide building blocks. Kornberg's Stanford group then synthesized infectious viral DNA. Arthur Kornberg 1970 ONCOGENES Peter Duesberg and Peter Vogt, virologists at UCSF, discovered the first oncogene in a virus. This SRC gene has since been implicated in many human cancers.

43 Biotechnology Time Lines 1972 FIRST RECOMBINANT DNA MOLECULE Paul Berg isolated and employed a restriction enzyme to cut DNA. Berg used ligase to paste two DNA strands together to form a hybrid circular molecule. This was the first recombinant DNA molecule. Paul Bergrecombinant DNA 1972 NIH GUIDELINES FOR RECOMBINANT DNA In a letter to Science, Stanford biochemist Paul Berg and others called for the National Institutes of Health to enact guidelines for DNA splicing.. Their concerns eventually led to the 1975 Asilomar Conference.

44 Biotechnology Time Lines 1973 AMES TEST Bruce Ames, a biochemist at UC Berkeley, developed a test to identify chemicals that damage DNA. The Ames Test becomes a widely used method to identify carcinogenic substances-carcinogen RECOMBINANT DNA MORITORIUM A moratorium on recombinant DNA experiments was called for at an international meeting at Asilomar, California, where scientists urged the government to adopt guidelines regulating recombinant DNA experimentation. The scientists insisted on the development of "safe" bacteria and plasmids that could not escape from the laboratory

45 Biotechnology Time Lines 1976 MORE ABOUT ONCOGENES J. Michael Bishop and Harold Varmus, virologists at UCSF, showed that oncogenes appear on animal chromosomes, and alterations in their structure or expression can result in cancerous growth. J. Michael BishopHarold Varmus 1976 RELEASE OF NIH GUIDELINES The NIH released the first guidelines for recombinant DNA experimentation. The guidelines restricted many categories of experiments.

46 Biotechnology Time Lines Present: The Dawn of Biotech Genetic engineering became a reality when a man-made gene was used to manufacture a human protein in a bacteria for the first time. Biotech companies and universities were off to the races, and the world would never be the same again. In 1978, in the laboratory of Herbert Boyer at the University of California at San Francisco, a synthetic version of the human insulin gene was constructed and inserted into the bacterium Escheria coli. Since that key moment, the trickle of biotechnological developments has become a torrent of diagnostic and therapeutic tools, accompanied by ever faster and more powerful DNA sequencing and cloning techniques.Herbert Boyer

47 Biotechnology Time Lines 1977 Genentech, Inc., reports the production of the first human protein manufactured in a bacteria: somatostatin, a human growth hormone-releasing inhibitory factor. For the first time, a synthetic, recombinant gene was used to clone a protein. Many consider this to be the advent of the Age of Biotechnology. Genentech 1978 RECOMBINANT INSULIN Genentech, Inc. and The City of Hope National Medical Center announced the successful laboratory production of human insulin using recombinant DNA technology.

48 Biotechnology Time Lines 1980 PATENTS ALLOWED The U.S. Supreme Court ruled in that genetically altered life forms can be patented a Supreme Court decision in 1980 allowed the Exxon oil company to patent an oil-eating microorganism. Kary Mullis and others at Cetus Corporation in Berkeley, California, invented a technique for multiplying DNA sequences in vitro by, the polymerase chain reaction (PCR). Kary Mullispolymerase chain reaction

49 Biotechnology Time Lines 1982 Genentech, Inc. received approval from the Food and Drug Administration to market genetically engineered human insulin The U.S. Food and Drug Administration approves the first genetically engineered drug, a form of human insulin produced by bacteria. Michael Smith at the University of British Columbia, Vancouver, developed a procedure for making precise amino acid changes anywhere in a protein. SITE DIRECTED MUTAGENESIS.

50 Biotechnology Time Lines 1983 Eli Lilly received a license to make insulin Genetic fingerprinting enters the court room. Cal Bio cloned the gene that encodes human lung surfactant protein, a major step toward reducing a premature birth complication. Genetically engineered plants resistant to insects, viruses, and bacteria were field tested for the first time. The NIH approved guidelines for performing experiments in gene therapy on humans.

51 Biotechnology Time Lines 1986 The FDA granted a license for the first recombinant vaccine (for hepatitis) to Chiron Corp. The EPA approved the release of the first genetically engineered crop, gene-altered tobacco plants Calgene, Inc. received a patent for tomato polygalacturonase DNA sequence, used to produce an antisense RNA sequence that can extend the shelf-life of fruit.

52 Biotechnology Time Lines 1988 Harvard molecular geneticists Philip Leder and Timothy Stewart awarded the first patent for a genetically altered animal, a mouse that is highly susceptible to breast cancer UCSF and Stanford University were issued their 100th recombinant DNA patent license. By the end of fiscal 1991, both campuses had earned $40 million from the patent. PATENTS AND MONEY

53 Biotechnology Time Lines 1990 The first gene therapy takes place, on a four-year-old girl with an immune-system disorder called ADA deficiency. The therapy appeared to work, but set off a fury of discussion of ethics both in academia and in the media. The Human Genome Project, the international effort to map all of the genes in the human body, was launched. Estimated cost: $13 billion Formal launch of the international Human Genome Project.Human Genome Project Publication of Michael Crichton's novel Jurassic Park, in which bioengineered dinosaurs roam a paleontological theme park; the experiment goes awry, with deadly results.Jurassic Park

54 Biotechnology Time Lines 1992 The U.S. Army begins collecting blood and tissue samples from all new recruits as part of a "genetic dog tag" program aimed at better identification of soldiers killed in combat Kary Mullis won the Nobel Prize in Chemistry for inventing the technology of polymerase chain reaction (PCR).PCR

55 Biotechnology Time Lines 1994 The first genetically engineered food product, the Flavr Savr tomato, gained FDA approval.Flavr Savr The first crude but thorough linkage map of the human genome appears.(See Science, v.265, Sep.30, '94, for the full color pull-out) A new coalition of mainstream religions launched a campaign seeking to overturn current laws allowing the patenting of genes used for medical and research applications. The group also includes Jeremy Rifkin, the controversial and outspoken critic of the biotechnology industry. SHOULD PATENTS BE ALLOWED? new coalition Jeremy Rifkin

56 Biotechnology Time Lines 1996 The discovery of a gene associated with Parkinson's disease provides an important new avenue of research into the cause and potential treatment of the debilitating neurological ailment.gene associated with Parkinson's disease A new inexpensive diagnostic biosensor test for the first time allow instantaneous detection of the toxic strain of E. coli E. coli strain 0157:H7, the bacteria responsible for several recent food-poisoning outbreaks. CAN IT BE DONE FOR ANTRAX OR OTHER BIOTERRORISM AGENTS?biosensor Surveys indicate the public regards research into the workings of the human genome and gene therapy with a combination of fear and mistrustfear and mistrust

57 Biotechnology Time Lines 1997 Researchers at Scotland's Roslin Institute report that they have cloned a sheep--named Dolly-- from the cell of an adult ewe. Dolly the first sheep cloned by nuclear transfer technology bearing a human gene appears later. Nuclear transfer involves transferring the complete genetic material (the DNA contained in a nucleus) from one cell into an unfertilized egg cell whose own nucleus has been removed.Dolly.

58 Biotechnology Time Lines 1998 Two research teams succeed in growing embryonic stem cells, the long sought grail of molecular biology.embryonic stem cells Scientists at Japan's Kinki University clone eight identical calves using cells taken from a single adult cow.clone eight identical calves A rough draft of the human genome map is produced, showing the locations of more than 30,000 genes.genome map

59 Biotechnology Time Lines 1999 MAD COW DISEASEA new medical diagnostic test for the first time allow quick identification of BSE/CJD a rare but devastating form of neurologic disease transmitted from cattle to humans.BSE/CJD

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61 Application of Biotechnology Its use and application ranges from fields like agriculture to industry (food, pharmaceutical, chemical, bioproducts, textiles etc.), medicine, nutrition, environmental conservation, animal sciences etc. making it one of the fastest growing fields. The work is generally carried out in the laboratories, as it is a scientific research oriented field.

62 Current Applications of Biotech Before going in-depth with a few areas of biotech research, it is important to gain a brief understanding of the many applications of biotechnological advancements. Microbial Agricultural Animal Forensic Environmental Aquatic Medical

63 RED BIOTECHNOLOGY: Medicine and pharmaceutics GREEN BIOTECNOLOGY: Agriculture and food WHITE BIOTECNOLOGY: Industrial proccesses BLUE BIOTECNOLOGY: Environment Biotecnology Application

64 Biotech Applications Biopolymers and Medical Devices- natural substances useful as medical devices  hyaluronate- an elastic, plastic like substance used to treat arthritis, prevent postsurgical scarring in cataract surgery, used for drug delivery  adhesive substances to replace stitches Designing Drugs – using computer modeling to design drugs without the lab- protein structure

65 Biotech Applications Replacement Therapies- lack of production of normal substances  Factor VIII- missing in hemophilia  Insulin Use of Transgenic Animals and Plants

66 Biotech Applications Gene Therapy – replace defective genes with functional ones  ADA (adenosine deaminase) deficiency  cystic fibrosis Immunosuppressive Therapies – used to inhibit rejection (organ transplants) Cancer Therapies -one method is antisense technology Vaccines – biggest break through in biotechnology- prevention of disease

67 Products of Modern Biotechnology There are a wide variety of products that the biotechnology field has produced. More than 65% of biotech companies in the U.S. are involved in pharmaceutical production (relating to drugs developed for medical use) Genentech developed Humulin (human insulin) to treat diabetes. It was the first biotech drug to be FDA approved.

68 Products of Modern Biotechnology There are more than 80 biotech drugs, vaccines, and diagnostics with more than 400 biotech medicines in development targeting over 2oo diseases! Nearly 1/2 of new drugs target cancer

69 Top 10 Selling Biotech Drugs DrugDeveloperFunction BetaseronChiron/BerlexMultiple sclerosis CeredaseGenzymeGaucher’s disease Engerix BGenentechHepatitis B vaccine EpiverGlaxoSmithKleinAnti-HIV EpogenAmgenRed blood cell enhancement GenotropinGenentechGrowth failure HumulinGenentechDiabetes IntronBiogenCancer & viral infections NeupogenAmgenNeutropenia reduction ProcritAmgenPlatelet enhancement

70 Biotech Treatments In the near future, it may be commonplace for treatments to include the use of gene therapy (attempt to replace “ defective ” gene with “ normal ” gene) and tissue engineering (designing & growing tissues for use in regenerative medicines). 1st Genetically Modified Organism (GMO) to produce human protein was E. coli (pictured right) that was given DNA to produce somatostatin (hGH - human growth hormone )

71 tPA One of the first genetically engineered (GE) products sold was tissue plasminogen activator (tPA) tPA is a blood clot dissolving enzyme used immediately after a heart attack or stroke to clear blocked vessels

72 Other Biotech Products Other biotech products include proteins in: home pregnancy tests (monoclonal antibodies) frost-resistant strawberry plants Although many are focused on medical and agricultural applications, some are for our own fashion interests (specialty apparel)!

73 Genes for Jeans? Stonewashed jeans use genetically engineered enzymes (amylase & cellulase) to create a faded look Originally, pumice stones were used (jeans washed with the stones) This method damaged the machines

74 Microbial Applications Bacteria & yeast are the most frequently used microbes Better enzymes and organisms for making foods, simplifying manufacture and production processes, and making decontamination processes for industrial waste product removal more efficient. Microbes used to clone and produce batch amounts of important proteins

75 Agricultural Applications Agricultural Biotechnology is estimated to be $6 billion market (2005), including applications such as: Pest-resistant plants Higher protein & vitamin content in foods Drugs developed and grown as plant products Drought-resistant, cold-tolerant, and higher-yielding crops

76 Plant Advantage The Ag-Biotech field boasts about the plant advantage over microbial biotech. Plant advantage refers to the fact that the cost of producing plant material with recombinant proteins is often significantly lower than bacteria Also, the Ag biotech may combine with medical biotech in order to produce drugs with molecular pharming

77 Molecular Pharming Molecular pharming is the use of genetically modified plants (or animals) as a source of pharmaceutical products. These are usually recombinant proteins with a therapeutic value. This is an emerging but very challenging field that requires: manipulation (at the genetic engineering level) of protein glycosylation (addition of polysaccharide chain) subcellular protein targeting in plant cells

78 Animal Applications Animals can be used as bioreactors! Many human therapeutic proteins are needed in massive quantities (>100s of kgs), so scientists create female transgenic animals to express therapeutic proteins in milk. Goats, cattle, sheep, & chickens are sources of antibodies (protective proteins that recognize & destroy foreign material) Transgenic refers to containing genes from another source

79 Dolly In 1996, Dolly sheep became the first cloned animal created by somatic cell nuclear transfer process. Born: July 5, 1996 Announced: February 22, 1997 Died: February 14, 2003 Dolly was cloned from a cell taken from a six-year-old ewe She became the center of much controversy that still exists today

80 Dolly gave birth to four lambs Dolly the sheep, the first mammal to be cloned from an adult cell

81 Cloned kitty The first two cats cloned by chromatin transfer

82 Human Clone Britain grants embryo cloning patent and became the first country in the world to grant a patent covering cloned early-stage human embryos. The decision ignited new controversy among biotechnology critics even though the Geron Corporation, the company licensed to use the patent, has no intention of creating cloned humans.

83 Cloning in the public eye: myths and misconceptions!!! Bush says cloning human embryos is 'morally wrong.'

84 Knock Outs Basic research in biotech uses knock-out experiments, which are very helpful for learning about the function of a gene. A knock-out is created when an active gene is replaced with DNA that has no functional information. Without the gene present, it may be possible to determine how the gene affects the organism (its function)

85 Aquatic Applications Aquaculture is a common aquatic application of biotech. Aquaculture is the process of raising finfish or shellfish in controlled conditions for food sources. Products include: transgenic salmon (increased growth rates) disease-resistant oysters vaccines against viruses that infect aquatic species Overall, aquatic organisms are thought to be rich & valuable sources for new genes, proteins, & metabolic processes.

86 Medical Applications Medical applications of biotech include preventative, diagnostic, and treatment. The Human Genome Project is very useful within this field. Gene therapy and stem cell technologies are two up-and- coming fields within the medical area of biotech. Stem cell technologies include immature cells that have the potential to develop and specialize into a variety of other cell types.

87 Stem Cells Different chemicals can coax them to develop into different cell types. Newest, most promising area Most controversial

88 Forensic Applications DNA fingerprinting is the classic example of a forensic application. It is used most commonly for law enforcement and crime scene investigation (CSI). It was first used in 1987 to convict a rapist in England. Other applications of DNA fingerprinting include: identifying human remains paternity tests endangered species (reduces poaching) epidemiology (spread of disease )

89 Environmental Applications The major environmental use is for bioremediation. Bioremediation is the use of biotech to process or degrade a variety of natural and manmade products, especially those contributing to pollution Therefore, cleaning up environmental hazards produced by industrial progress is a major application of this type of biotechnology. There is a strong tie to microbial biotech (since many microbes are helpful for this area).

90 Bioremediation Bioremediation can be defined as any process that uses microorganisms or their enzymes to return the environment altered by contaminants to its original condition.microorganisms enzymesenvironmentcontaminants

91 ENVIRONMENT Environmental biotechnology has become another area of extensive work due to the dangers brought about by increasing levels of environmental pollution. A lot of hard work is being done to protect our environment. In this field, the job of a biotechnologist spans from checking industrial air pollution levels, treatment of industrial waste to recycling of sewage sludge.

92 Oil Spill In the 1970s, the first U.S. GMO patent was granted to a scientist for a strain of bacteria capable of degrading components in crude oil. In 1989, the Exxon Valdez oil spill in Alaska used Pseudomonas species (oil- degrading bacteria) to clean up the spill It was 3x faster & without increased environmental effects

93 Waste Management  Environmental Pollution is a major problem  Landfills are becoming full  Old dump sites are creating problems  Waste is piling up  Sewage and chemical disposal is a constant problem

94 Waste Management  Genetically altered bacteria are used to feed on oil slicks and spills  Bacteria are being developed to decompose or deactivate dioxin, PCBs, insecticides, herbicides, and other chemicals  Bacteria are under development to convert solid wastes into sugars and fuel

95 Biomaterials Biotechnology is the technology connected with plant, animal, and human life. For example, knowledge about the joints, muscles, and nerve endings in our bodies can be combined with engineering to develop an artificial hand. These human-made materials designed to be placed within the human body are called biomaterials.

96 Transplant Organs for Humans Inadequate supplies of human organs for transplantation result in more than 25,000 deaths annually. Researchers are studying ways to develop organs in animals suitable to be transplanted into humans until human organs become available. Especially promising are pig organs!!!

97 Photo of mouse growing a "human ear" - a shape made of cartilage

98 Biohand: knowledge from the joints, muscles, and nerve

99 Genetically Modified Food Can animal genes be jammed into plants? Would tomatoes with catfish genes taste fishy? Have you ever eaten a genetically modified food? The answers are: “ yes ”, “ no ” and almost definitely “ yes ” Despite dire warnings about "Frankenfoods “, there have been no reports of illness from these products of biotechnology.

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102 GM potato 'could improve child health'

103 Approved Biotech Products 1982: FDA approves genetically engineered human insulin 1986: Orthoclone OKT3 (Muromonab-CD3) approved for reversal of kidney transplant rejection. 1986: first recombinant vaccine approved- hepatitis 1987: Genentech gets approval for rt-PA (tissue plasminogen activatior) for heart attacks

104 Approved Biotech Products 1990: Actimmune (interferon 1b) approved for chronic granulomatous disease Adagen (adenosine deaminase) approved for severe combined immunodeficiency disease. 1994: first genetically engineered food the Flavr Savr tomato is approved. 1994: Genentech ’ s Nutropin is approved (growth hormone deficiency)

105 Approved Biotech Products 1994: Centocor ’ s ReoPro approved (for patients undergoing balloon angioplasty) Genzymes Ceredase/Cerezyme approved for Gaucher ’ s Disease (inherited metabolic disease) Recombinant GM-CSF approved (chemotherapy induced neutropenia)  1998: Centocor ’ s RemicadeTM approved (monoclonal antibody for Crohn ’ s disease)

106 Focus on “ Famous ” Biotech Product: Insulin Insulin: Insulin is a hormone, and therefore, a protein. Insulin was the first hormone identified (late 1920's) which won the doctor and medical student who discovered it the Nobel Prize (Banting and Best). They discovered insulin by tying a string around the pancreatic duct of several dogs. Note that there are other hormones produced by different types of cells within pancreatic islets (glucagon, somatostatin, etc) but insulin is produced in far greater amounts under normal conditions making the simple approach used by Banting and Best quite successful.glucagon, somatostatin, etc

107 Properties of Insulin Insulin is secreted by groups of cells within the pancreas called islet cells. The pancreas is an organ that sits behind the stomach and has many functions in addition to insulin production. The pancreas also produces digestive enzymes and other. Without insulin, you can eat lots of food and actually be in a state of starvation since many of our cells cannot access the calories contained in the glucose very well without the action of insulin.

108 Insulin (cont ’ d) The first successful insulin preparations came from cows (and later pigs). The pancreatic islets and the insulin protein contained within them were isolated from animals slaughtered for food in a similar but more complex fashion than was used by our doctor and med-student duo.

109 Wockhardt launches indigenous human recombinant insulin Mr H. Khorakiwala, Chairman, Wockhardt Ltd, with Dr M. Sahib, Director, Genomics & Biotech Research, at the launch of human recombinant insulin `Wosulin' in Mumbai

110 Thank you

111 Biotech Applications Diagnostics  Antibodies  Biosensors  PCR Therapeutics  Natural Products  Foxglove:  digitalis: heart conditions Yew tree- cancer agent (taxol) breast and ovarian cancers Endogenous Therapeutic agents – proteins produced by the body that can be replicated by genetically engineered: tPA – tissue plasminogen factor (dissolves blood clots)

112 Approved Biotech Products 1938: Howard Florey/Ernst Chain, Oxford U., England isolated penicillin : Large scale production of penicillin : Cortisone first manufactured in large amounts 1977: Genentech produced somatostatin (human growth hormone-releasing inhibitory factor), manufactured in bacteria. First time a recombinant gene was used to clone a protein. 1978: Harvard researchers produced rat insulin by recombinant DNA.


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