Biotechnology - PowerPoint Presentation

Slide1

BiotechnologySlide2

Learning Goals

Describe the science of Biotechnology and its product domains

List the steps to producing a GMO through use of

rDNA

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 forSlide3

Learning Goals cont.

Apply the strategy for values clarification to bioethical issues

Identify products made through biotechnology

Identify products made through biotechnologySlide4

Vocabulary

Insulin

Biotechnology

DNA

rDNA

technology

PCR

Cloning

FermentationDiabetesAntibioticsRestriction enzymeDNA ligaserDNA

Proteases

Antibodies

Pharmaceutical

R&D

Pure Science

NIH

CDC

DNA Fingerprinting

Microbial agents

Virus

Applied science Slide5

Vocab cont.

Moral

GMO

E. coli

Plasmid

t-PA

Efficacy

Clinical trials

FDACystic fibrosisBiochemistryMolecular biologyGenetics ethics

Therapeutic

EPA

USDA

Data

Hypothesis

Variable

Control

Positive control

Negative control

Concentration

Journals

Genome

bioethicsSlide6

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

Scientists recently learned to manipulate not only whole organisms, but also the molecules, cells, tissues, and organs of which they are builtSlide7

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 sugarSlide8

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 Slide9

Biotechnology: Any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use

Past

Present

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 (

rDNA

) technology

PCR

Cloning

ProteomicsSlide10

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 familySlide11

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 virusesSlide12

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 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!Slide13
Slide14

Importance of rDNA

technology

Advances in

rDNA

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 proteinsAntibodies: proteins normally developed by the immune system for recognizing specific molecules called antigens on infectious agents and so limiting their infectious abilitySlide15

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 soSlide16

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.Slide17

Biotechnology Workplace

Biotechnology is practiced in a variety of settings , and in general the setting determine the major emphasis

Private companies: focus is making money so they tend to emphasize the development and manufacture of products that will yield high profit margins

The do have a Research & Development department, but they will only fund research that has potential for financial pay off

Universities & Government agencies focus on research

More involved in Pure science

The main difference between companies and university or government labs is that companies must provide a product or service that results in earnings, a nonprofit research facility does notSlide18

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 onSlide19

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 1.13Slide20

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 conditionsSlide21

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 healthSlide22

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

rDNA

rDNA

can be inserted into cells giving these cells a new genotype phenotype

Organisms containing these modified cells are called Genetically Modified Organisms (GMOs)Slide23

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

) bacteriaE. coli bacteria are the work horse of molecular biology, no other bacteria is used as frequently as this oneSlide24

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 useSlide25

Tissue Plasminogen Activator

One of the 1

st

genetically engineered products to be sold was t-PA

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) cellsSlide26

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 1.18Slide27

Biotechnology & Society

Applications of

rDNA

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

rDNA techniques undergoing clinical trials Strawberry plants that can grow in freezing weatherSlide28
Slide29

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 accordinglySlide30

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 introSlide31

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 HER2

The decision to administer

Herceptin as part of the chemotherapy regimen is based on a single geneSlide32

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 nowSlide33

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”Slide34

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 PlanSlide35

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 1.21Slide36

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 the FDA before being marketed

Example:

Pulmozyme

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 30Pulmozyme reduces the amount of mucus produced

In 2003

Pulmozyme

had sales of $167million Slide37

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 animalsSlide38

Biotechnology with a Conscience

Being able to distinguish between right and wrong and to make decisions based on that knowledge is considered having good morals

Problem: not all people have the same list of right and wrong, and so we often have different morals

Example: Vegans don’t eat or use any animal products, and many consider it immoral to do so

Fact: most people eat and use animal products on a daily basis without any qualms

Also Fact: those same people that don’t have any problem eating meat and using animal products would be horrified to think of going home and cooking their pet dog Sparky or their new kitten Mittens for dinnerSlide39

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 technologySlide40

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 GodSlide41

Technology is Not Evil

New technologies generate ethical questions

The positions one takes on ethical issues 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”

But sometimes people are!Slide42

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 itSlide43

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 researchSlide44

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 dataFormulate conclusions based on experimental data and error analysis Slide45

Control Groups

Positive Controls

Negative controls

A group that will give a predictable positive result

A group that will give a predictable negative resultSlide46

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 ErrorsIdentify the sources of expreimental design errors that would or could lead to false or misleading data, and explain possible implications from such errorsPA: 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 hereSlide47

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 or production instruments, reagents, or dataSlide48

Review: List the steps to producing a GMO through use of rDNA

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

rDNA will be used by the host organism to produce the new protein of interestSlide49

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 TrialsFDA approvalSlide50

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 animalsSlide51

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 aspectsRank solutions from best to worstDecide and defend your positionSlide52

Identify products made through biotechnology

T-PA

Human insulin

Strawberry plants that can grow in freezing weather

Pulmozyme