BIOTECHNOLOGY Learning Goals Describe the science of Biotechnology

BIOTECHNOLOGY

Learning Goals � Describe the science of Biotechnology and its product domains � List the steps to producing a GMO through use of r. DNA � Outline the steps of producing and delivering a product made through recombinant DNA technology � Identify the important labs and agencies in the US, and what they’re mainly responsible for

Learning Goals cont. � Apply the strategy for values clarification to bioethical issues � Identify products made through biotechnology

Vocabulary � � � Insulin Biotechnology DNA r. DNA technology PCR Cloning Fermentation Diabetes Antibiotics Restriction enzyme DNA ligase r. DNA � � � Proteases Antibodies Pharmaceutical R&D Pure Science NIH CDC DNA Fingerprinting Microbial agents Virus Applied science

Vocab cont. � � � � Moral GMO E. coli Plasmid t-PA Efficacy Clinical trials FDA Cystic fibrosis Biochemistry Molecular biology Genetics ethics � � � � Therapeutic EPA USDA Data Hypothesis Variable Control Positive control Negative control Concentration Journals Genome bioethics

Defining Biotechnology � We have been manipulating entire organisms for thousands of years through selective breeding to produce plant and animal products that are � Bigger � Healthier � More productive � Have novel phenotypes q Scientists recently learned to manipulate not only whole organisms, but also the molecules, cells, tissues, and organs of which they are built

Example: Human Insulin � � � Insulin is a protein produced in healthy individuals that facilitates the uptake of sugar from the blood stream into specific tissues In some diabetic patients, the ability to produce insulin is impaired In the past diabetic patients had to take injections of Bovine (cow) insulin to regulate their blood sugar

Example: Human Insulin � � Problem: getting insulin from cows is not very efficient, it’s time consuming and expensive, and can lead to improper immune stimulation of the recipients Solution: Biotechnology � We have learned to manufacture large amounts of specific molecules, such as human insulin in bacterial cells Cost effective Convenient Fast

Biotechnology: Any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use PRESENT PAST � � Selective breeding Fermentation: a process by which in an oxygen deprived environment a cell converts sugar into lactic acid or ethyl alcohol (ethanol) � � � DNA technology Recombinant DNA (r. DNA) technology PCR Cloning Proteomics

Modern Biotechnology � � Since biotechnology is constantly changing there is no one definitive definition In 2003 in an international treaty on biotechnology, modern biotechnology was defined as : � “the application of techniques that overcome natural physiological reproductive or recombination barriers and that are not techniques used in traditional breeding and selection “ including In vitro nucleic acid techniques The fusion of cells beyond taxonomic family

Restriction Enzymes � � � One major discovery that propelled the biotechnology science and industry was the discovery of restriction enzymes Restriction enzymes: enzymes that cut DNA at specific recognition nucleotide sequences known as restriction sites They were discovered in bacteria & are thought to have evolved to provide a defense mechanism against invading viruses

Recombinant DNA Technology � � Restriction enzymes allow us to cut and recombine DNA molecules, often from different species “cutting” � Reminder DNA (deoxyribonucleic acid) is a double stranded helical molecule that stores genetic information for the production of all of an organisms proteins � DNA Ligase: an enzyme that binds the disconnected strands of DNA together � � “pasting” If you put DNA into an organisms cell, that cell will produce the protein that DNA codes for!

Importance of r. DNA technology � � Advances in r. DNA technology allow biotechnologists to manufacture a wide variety of products either previously unavailable or that could only otherwise be made in small quantities Examples: � Insulin: for treatment of diabetes � Proteases: function in breaking down other proteins � Antibodies: proteins normally developed by the immune system for recognizing specific molecules called antigens on infectious agents and so limiting their infectious ability

Other Helpful Techniques � PCR: Polymerase Chain Reaction is a technique that involves copying short pieces of DNA and using them as templates to make millions of copies in a very short amount of time � Cloning: method of asexual reproduction that produces identical organisms � Done mainly in bacteria and some fungi � Only happens in animals if manipulated by scientists to do so

Cao Yilin, a plastic surgery expert in Shanghai, successfully reproduces a human ear on the body of a little white mouse on Friday, April 4, 1997 in Shanghai. Dolly, right, the first cloned sheep produced through nuclear transfer from differentiated adult sheep cells, and Polly, the world's first transgenic lamb, are in their pen at the Roslin Institute in Edinburgh, Scotland, in early December, 1997.

Biotechnology Workplace � Biotechnology is practiced in a variety of settings , and in general the setting determine the major emphasis The main difference between companies: focus is making money so they companies and university or tend to emphasize the development and government labs is that manufacture of products that companies will yield high profit must provide a product or service that margins earnings, a nonprofit department, The doresults have ain. Research & Development research facility doesthat nothas potential for but they will only fund research � Private financial pay off � Universities research & Government agencies focus on More involved in Pure science

Pure vs. Applied Science � Pure-science: Scientific research whose main purpose is to enrich the scientific knowledge base � This � is the type of science nonprofit labs focus on Applied Science: the practice of utilizing scientific knowledge for practical purposes, including the manufacture of a product � This is the type of research forprofit labs focus on

4 Main Categories of Biotechnology Companies � � Pharmaceutical Agricultural Industrial Research or production instruments, reagents, or data � Some companies sell their services rather than product � Example: 23 and Me � See figure 1. 13

R&D vs. QC � � Research & Development : refers to the early stages of product development that includes discovery of the structure and function of a potential product and the initial small scale production Quality Control is a measure of precision or how well the measurement system reproduces the same result over time and under varying operating conditions

Important Government Labs � National Institute of Health (NIH): the federal agency that funds and conducts biomedical research � Centers for Disease Control & Prevention (CDC): National research center for developing and applying disease prevention control, environmental health, and health promotion and educationactivities to promote public health

Bioengineered Products � � � By the 1970’s scientists had developed new methods including the use of restriction enzymes and DNA ligase to create new combinations of genetic information, or r. DNA can be inserted into cells giving these cells a new genotype phenotype Organisms containing these modified cells are called Genetically Modified Organisms (GMOs)

GMOs � � � GMO’s contain DNA from another organism and will produce the new proteins encoded in the acquired DNA The 1 st GMO’s to produce a human protein were Escherichia coli (E. coli) bacteria E. coli bacteria are the work horse of molecular biology, no other bacteria is used as frequently as this one

Escherichia coli � � Were given pieces of human DNA (genes) containing instructions to produce a human growth hormone called somatostatin The somatostatin gene was carried into the E. coli cells on tiny pieces of bacterial DNA called Plasmids � These recombinant DNA plasmids contained both bacterial and human DNA � The E. coli cells read the human DNA and produced the human protein somatostatin, which could then be harvested and purified for use

Tissue Plasminogen Activator � � One of the 1 st genetically engineered products to be sold was t-PA is a blood clot dissolving enzyme that can be used immediately after a heart attack to clear blocked blood vessels � The body produces only small amounts of t-PA � To produce enough t-PA for therapeutic use scientists genetically engineered mammalian cells using Chinese Hamster Ovary (CHO) cells

Chinese Hamster Ovary � � � Ovary cells are grown in culture and engineered with the gene that provides the instructions to make the t-PA enzyme By subjecting the cells to the right conditions, the CHO cells accept and incorporate the foreign DNA and begin the process of transcription and translation Large amounts of t-PA can then be extracted and purified � See figure 1. 18

Biotechnology & Society � Applications of r. DNA and genetic engineering have resulted in the creation of hundreds of Biotech companies , specializing in all kinds of GMOs and their protein products including � Proteins used in pregnancy tests � Enzymes that increase the amount of juice that can be extracted from an apple � Molecules used in vaccines In 2004 there were already over 370 drug products created from r. DNA techniques undergoing clinical trials � Strawberry plants that can grow in freezing weather

Biotechnology & Healthcare � Biotechnological advances will enable patients to submit tissue samples and receive a treatment regimen designed specifically for them. This is calle Personalized medicine, or pharmacogenomics � pharmacogenomics: pharmaceuticals based on genomics � Present research based on broad characteristics such as ethnicity, gender, and family history has already been performed and treatments have been modofied accordingly

The Human Genome Project � � The human genome project was an international effort to sequence and map all the DNA on the 23 pairs of human chromosomes We know the sequence of the human genome, but we still do not know � all of the sequences that code for proteins � For the ones we do know about, we don’t completely understand how, when, and where they are turned on or off � This is called differential gene expression � DNA intro

Biotechnology & Healthcare � � � As sequencing human genomes becomes more cost effective, there will come a time when drug regimen is determined by the genotype of the patient or the tumor being targeted One of the 1 st drugs to accomplish this is the drug Herceptin which will only target breast cancer cells that produce too much of the protein HER 2 The decision to administer Herceptin as part of the chemotherapy regimen is based on a single gene

Biotechnology & Healthcare � � Some drugs, like chemotherapeutic agents, have serious side effects because they treat the whole organism and not just the diseased tissue Advances in biotechnology have enabled the development of therapies that can be delivered directly to diseased tissue and greatly reduce side effects � Rexin-G: a drug that targets pancreatic cancer is delivered by a genetically engineered viral particle that is designed to seek out and kill metastasized cancer cells is in clinical trials now

How Companies Select Products for Manufacture � � � Research & Development No matter what the product, the goal is to make it and market it as quickly as possible The R&D phase for a new pharmaceutical often takes from 10 -15 years for a company to move a product through the “pipeline”

Product Pipeline � A drug must demonstrate “proof of concept” data in the research phase before being allowed to move to the development phase, including � Is it feasible to manufacture in amounts sufficient to treat people � What needs to be done to ensure safety � What characteristics indicate it will be effective � And unfortunately, what is the profit margin � This is accomplished with a Product development Plan

Product Development Plan � Does the product meet the following criteria � Does the product meet a critical need? � Is there a market large and wealthy enough to produce enough sales � Does preliminary data support that the product will work? � Can patent protection be secured � Can the company make a profit? � � Each product in a pipeline will be reviewed regularly and dropped from production if the answers to these questions are unsatisfactory See figure 1. 21

Product Pipeline � � Projects then enter clinical development Testing is done small scale before large scale production is pursued All pharmaceuticals have to undergo at least 3 rounds of strict clinical trials under the guidance of In 2003 Pulmozyme the FDA before being had marketed sales of Example: Pulmozyme $167 million is manufactured by Genentech Inc and is an enzyme used to treat patients with Cystic Fibrosis � People with CF experience a build up of mucus in the lungs and intestines and is often fatal by the age 30 � Pulmozyme reduces the amount of mucus produced

Regulations Governing Product Development � All biotechnology products have regulations governing their production in the product pipeline � Environmental Protection Agency (EPA) The federal agency that enforces environmental laws including the production and use of microorganisms, herbicides, pesticides, and GMOs � United States Department of Agriculture (USDA) Federal agency that regulates the use and production of plants, plant products, plant pests, veterinary supplies and medications, and genetically modified plants and animals

Biotechnology with a Conscience � � Being able to distinguish between right and wrong and to make decisions based on that Also Fact: those same knowledge is considered having goodhave morals people that don’t Problem: not all peopleany have the same problem eatinglist of right and wrong, andmeat so we often different and usinghave animal morals products would be to think Example: Vegans don’thorrified eat or use anyofanimal going home and to do products, and many consider it immoral cooking their pet dog so Sparky or their new Fact: most people eat and. Mittens use animal products kitten for dinner on a daily basis without any qualms

Bioethics � The study of moral standards and how they effect conduct is called ethics � Bioethics is a term that has been coined to describe the study of decision making as it applies to moral decisions that need to be made because of advances in biology, medicine, and technology

Bioethics � � Many of the new biotechnologies are controversial because they force people to think about what they believe is right or wrong Examples: � Harvesting and manipulating embryonic stem cells It kills the human embryo � Genetically modifying food Are they really safe to eat � Prenatal genetic testing/ engineering Playing God

Technology is Not Evil � � � New technologies generate ethical questions The. But positions one takes on ethical issues sometimes people are! are based on how you feel about the knowledge and understanding you have of a subject There is often not a clear right or wrong answer Most of the time you have to make a choice between the lesser of 2 evils So how do we decide what’s “right”

Strategy for Values Clarification � � � Identify and understand the problem or issue. List all the possible solutions to the issue Identify the pros and cons of each of the solutions � Consider legal, financial, medical, personal, social, and environmental aspects � � Based on the pros and cons for each solution, rank all the solutions from best to worst Decide if the problem is important enough to take a position. If it is, decide your position and be prepared to describe and defend it

Doing Biotechnology: Scientific Methodology in a Research Facility � � � There is no 1 correct scientific method The scientific method is really just a series of common sense steps and techniques that scientists use to solve a problem or answer a question There are several practices used by most scientists when conducting experimental research

Scientific Methodology � � � State a testable scientific question or problem based on some information or observation Develop a testable hypothesis Plan a valid experiment � Multiple replicates � Control groups � � Conduct the outlined experiment and collect and organize data Formulate conclusions based on experimental data and error analysis

Control Groups POSITIVE CONTROLS � A group that will give a predictable positive result NEGATIVE CONTROLS � A group that will give a predictable negative result

Writing Experimental Conclusions � � � A good approach to writing an experimental conclusion is to use the REE/PE/PA method REE: Results with Evidence and Explanation PE: Possible Errors � Identify the sources of expreimental design errors that would or could lead to false or misleading data, and explain possible implications from such errors � PA: Practical Applications: � discuss the meaning or value of experimental results in the short and long term. � Identify how the findings could be valuable clinically, to the company, or to the community � Identify the direction the experiment will go take from here

Review: Describe the science of Biotechnology and its product domains � � Biotechnology: The study and manipulation of living things or their component molecules, cells, tissues, or organs Product Domains � Pharmaceutical � Agricultural � Industrial � Research data or production instruments, reagents, or

Review: List the steps to producing a GMO through use of r. DNA � � � Identify a gene of interest Treat gene of interest and plasmid vectors with a restriction enzyme Use DNA ligase to glue gene of interest to plasmid Insert plasmid into host organism (plant, animal, or bacteria) through various mechanisms The r. DNA will be used by the host organism to produce the new protein of interest

Review: Outline the steps of producing and delivering a product made through recombinant DNA technology � � Come up with an Idea Research and Development � Proof of Concept through a Product Development Plan Will it work Is there a market for it Can we make it in sufficient quantities � Large Scale Production � Clinical Trials � FDA approval

Review: Identify the important labs and agencies in the US, and what they’re mainly responsible for � � NIH: the federal agency that funds and conducts biomedical research CDC: National research center for developing and applying disease prevention control, environmental health, and health promotion and education activities to promote public health EPA: The federal agency that enforces environmental laws including the production and use of microorganisms, herbicides, pesticides, and GMOs USDA: Federal agency that regulates the use and production of plants, plant products, plant pests, veterinary supplies and medications, and genetically modified plants and animals

Review: Apply the strategy for values clarification to bioethical issues � � � Identify /understand the issue List all the possible solutions Identify the pros and cons of each considering legal � financial, � Medica � personal � Social � environmental aspects � � � Rank solutions from best to worst Decide and defend your position

Identify products made through biotechnology � � T-PA Human insulin Strawberry plants that can grow in freezing weather Pulmozyme