BIOTECHNOLOGY IN MEDICAL INDUSTRY

Prepared by Hilal MARANGOZ Sevgi DOAN

What is medical biotechnology ?

Medical biotechnology is the use of living cells and cell materials to research and produce pharmaceutical and diagnostic products that help treat and prevent human diseases.

APPLICATION AREAS
Drug Production
Penicillin Insulin (DNA recombinant technology)

Artificial organs Gene therapy Vaccines Diagnostics Pharmacogenetics Biomedical Appications

Medical history
The role of bacteria in human diseases have been found by Robert Koch in 1877.

Discovery of animal insulin in 1921 by Frederick Grant Banting. Finding penicillin and using in treatment of diseases as an antibiotics is realized by Alexander Fleming in 1928.

Discovery of human insulin in 1978 by Genentechs scientists. Syrups and polysaccharides was produced within the next 30 years.

Antibiotics
Antibiotics are chemical substances produced by microorganisms to kill other microorganisms. There are two types antibiotcs which are biocidial and biostatic. They are used in the treatment of infectious diseases.

Through discovery of penicillin

Inflamed wounds were treated with moldy soybean in 600 BC in China.

Sumerian and Egyptians were used as an antibiotics produced beer from stale bread.

Wounds were treated with soil in Mesopotamio.

The story of discovery of penicillin by chance

In 1928, Alexander Fleming ,who was bacteriologist , augmented staphylococci in culture.
fungus

penicillin

Penicillin...
Penicillin , which is produced by fungus named as penicillum notatum , is bactericidial substance.

Penicillin have been purificated and used as a medicine by Florey and Chain in 1938 In 1945, Production of which reached to treat 1 milion patients.

Penicillin
All penicillin like antibiotics ,Inhibits enzymes involved in synthesis of peptidoglycan for bacterial cell wall, causing cell lysis.

They do not affect with the synthesis of other intracellular components.

These antibiotics do not affect human cells because human cells do not have cell walls.

Spectrum of Activity
Penicillins are active in Gram positive bacteria. Some members (e.g. amoxicillin) are also effective in Gram negative bacteria.

PRODUCTION OF PENICILLIN
Penicillin was produced by an aerobic fermentation.

It is fermented in a batch culture .

Primary and Secondary Metabolites

Primary metabolites are produced during active cell growth, and secondary metabolites are produced near the onset of stationary phase.

Commercial Production Of Penicillin

Like all antibiotics, penicillin is a secondary metabolite, so is only produced in the stationary phase.

INDUSTRIAL PRODUCTION OF ANTIBIOTIC- PENICILLIN

The industrial production of penicillin is broadly classified in to two processes namely,

Upstream processing Downstream processing

UPSTREAM PROCESSING
Upstream processing includes any technology that leads to the synthesis of a product. Upstream includes the exploration, development and production.

DOWNSTREAM PROCESSING

The extraction and purification of a biotechnological product from fermentation is referred to as downstream processing.

For penicillin

UPSTREAM PROCESSING

The medium is designed to provide the organism with all the nutrients that it requires.

RAW MATERIALS
CARBON SOURCES: Lactose acts as a very satisfactory carbon compound, provided that is used in a concentration of 6%. Others such as glucose & sucrose may be used.

NITROGEN SOURCES:

Corn steep liquor (CSL) Ammonium sulphate and ammonium acetate can be used as nitrogenous sources.

MINERAL SOURCES: Elements namely potassium, phosphorus, magnesium, sulphur, zinc and copper are essential for penicillin production. Some of these are applied by corn steep liquor.

STAGES IN DOWNSTREAM PROCESSING

Removal of cells The first step in product recovery is the separation of whole cells and other insoluble ingredients from the culture by technique such as filtration and centrifugation.

FERMENTATION PROCESS
Penicillin G, one of the most active and widely used forms, is manufactured commercially using Penicillium chrysogenum.

The process is carried out in stainless steel fermenters of l0000 dm3 capacity. The fermenter is steam sterilized and loaded with sterilized growth medium (corn steep liquor) containing lactose, amino acids, mineral salts and other substances.

FERMENTATION PROCESS
The pH requires adjustment from time to time, to neutralize ammonia produced by the fungus. Temperature is set at first to give the maximum growth rate and then altered to favor penicillin synthesis. The fermenter is continuously stirred and sterile air blown in. An external cooling jacket is used for temperature control.

FERMENTATION PROCESS
Penicillin is a secondary metabolite, produced in large quantities only towards the end of the growth period of the fungus. Therefore it is essential for all of the mycelium to reach peak growth at the same time. This is why batch fermentation, rather than a continuous process, is appropriate for penicillin manufacture. After about 200 hours, the broth is filtered. Penicillin passes through in the filtrate which is further processed to crystallize the product. After the seperation operation , penicillin can be modified by chemical subtances and semi-

The main stages of Penicillin production are:

FURTHER PROCESSING
For parental use, the antibiotic is packed in sterile vials as a powder or suspension. For oral use, it is tabletted usually now with a film coating. Searching tests (ex: for purity, potency) are performed on the appreciable number of random samples of the finished product. It must satisfy fully all the strict government

Production Ratio
When penicillin was first made using the fungus Penicilium notatum, the process made 1 mg dm-3. Today, using a different species (P.chrysogenum) and a better extraction procedures the yield is 50 g dm-3.

Usage of Antibiotics
Generally used to treat diseases caused by bacteria infection.
Used to prevent reproduction or to kill bacteria. Antibiotics , which are drug , are given at most by doctors. In recent years, widespread and uncontrolled use of antibiotics has caused to develop resistance to antibiotics of bacteria.

AUGMENTIN
In Turkey , Most used antibiotic is Augmentin.

Generally used for upper respiratory tract infection. Price is 8TL. Produced by Glaxo SmithKline

Turkey is number one consumption of antibiotic in the world

Unconscious using and selling of antibiotics provides acquire resistance to antibiotics. These types usage is widely in Turkey.Therefore we use more affective and expensive antibiotic than european countries.

ARTIFICIAL ORGANS
An artificial organ is a man-made device that is implanted or integrated into a human to replace a natural organ.

Classical Approaches to Artificial Organs

Technically and mechanically complex products, based on rather simple concepts.
Heart valve Heart Kidney Lung Eye Kidney Liver Pancreas valve pump filter gas-supplier feedback cameras biofeedback active bioreactors active bioreactors

IN FUTURE

Need for Artificial Organs?

Waitinglist Organ Transplantation USA 1990: 20.481 Patients 06/2006: 92.265 Patients
13.000 12.000 11.000 10.000 9.000 8.000 7.000 6.000 5.000 4.000 3.000 2.000 1.000 0 UNOS, June 07 2006

Waitinglist Kidney-Tx Germany Kidney - Transplantation in Germany Need ca. 3.500 / Year 2516 in 2003 2478 in 2004

1979

'81

'83

'85

'87

'89

'91

'93

'95

'97

'99

2001

Year

The Artificial Heart

Two successful manufacturers: - 1. Syncardia (formerly Jarvik) temporary CardioWest Total Artificial

Heart

- Used as a bridge between hearts for patients on organ waiting lists. - more than 800 implants - 79% of patients survived to transplant

The Artificial Heart

2.

AbioCor replacement heart:

the Massachusetts based company AbioMed has produced the AbioCor artificial heart. It is fully implantable within a patient, meaning that no wires or tubes penetrate the skin, thus reducing the risk of infection. To date, 15 patients have been implanted with the AbioCor,

The Artificial Lung

Currently in the last stages of development, the MC3 Company has introduced the BioLung. The BioLung is designed to replace the gas exchange function of a persons native lungs during recovery from injury or illness, or until donor lungs are available for transplantation.

The Cochlear Implant

Frequently

called the bionic ear, a cochlear implant is a surgically implanted electronic device that provides sound to a person who is profoundly deaf or severely hard of hearing.

CENTRE OF EXCELLENCE (BIOMATEN) IN TURKEY

BIOMATEN was established in ODT with the support of DPT

In the BIOMATEN, particularly bone and cartilage, bone cement, dental materials will be produced.

The scientific infrastructure will be prepared necessary products for the treatment of

ARTIFICIAL HEART IN EGE UNIVERSITY

Up to now , 25 patients implantted with artificial heart. Artificial hearts are bought from abroad. An artificial heart would cost 60 thousand dollars. Now , main target is to produce artificial heat.

idem Artificial heart

20 April,20011 In Ege University Hospital, 150 kilograms artificial heart device implanted to idem, who is 8.5 kilograms in weight.

Pending donation organ, she must live with implantted heart for the survival of her.

Nanotube plates

GENE THERAPY

The main purpose of gene therapy , instead of missing or defective genes causing disease of cells , healthy copies of the placement of the cell. Drug is not for the control of genetic defect, Instead, the defective genetic structure of patient is tinkered.

How it works ?
The target cells become infected with the viral vector A vector delivers the therapeutic gene into a patients target cell.

 The vectors genetic material is inserted into the target cell

Functional proteins are created from the therapeutic gene causing the cell to return to a normal state.

Successful One Year Gene Therapy Trial For Parkinson's Disease

Neurologix a biotech company announced that they have successfully completed its landmark Phase I trial of gene therapy for Parkinson's Disease.

This was a 12 patient study with four patients in each of three dose escalating cohorts. All procedures were performed under local anesthesia and all 12 patients were discharged from the hospital within 48 hours of the procedure, and followed for 12 months. Primary outcomes of the study design, safety were successfully met. There were no adverse events reported relating to the treatment

 The gene transfer procedure utilized the AAV (adeno-associated virus) vector, a virus that has been used safely in a variety of clinical gene therapy trials, and the vehicle that will be used in all of the company's first generation products, including epilepsy and Huntington's disease. In its Parkinson's disease trial, Neurologix used its gene transfer technology.

MAIN TOPICS
Biologics Insulin and diabetes Recombinant DNA Technology Diagnostics Pharmacogenetics

DRUGS
A drug is a therapeuatic substance used to prevent ,manage or cure disease. The pharmaceutical industry traditionally manufactures synthesized drugs, such as aspirin. The biotechnology revolution brought about a new class of drug: the biologic.

BIOLOGICS
Biologics (biological drugs) are therapies derived from living organisms. Biologics can be composed of;
sugars, proteins, nucleic acids, cells and tissues.

BIOLOGICS
Biologics includes;
Gene therapy which is mentioned Recombinant therapeutic proteins,
Biological drugs,such as insulin,growth hormone etc. DNA vaccines

INSULIN
Insulin is a hormone secreted by pancreas. It plays a key role in managing our blood sugar. Insulin has two main roles in ; Regulate Sugar in Blood Stream Storage of excess glucose for energy Insulin regulates carbohydrate and fat metabolism in the body and takes up glucose from the blood and stores glucose as glycogen in the liver and muscle.

DIABETES
If your pancreas secretes little or no insulin

(type 1 diabetes) or your body produces too

little insulin or has become resistant to insulin's action (type 2 diabetes), the level of

sugar in your bloodstream increases. This is

because it's unable to enter cells. High blood sugar can lead to complications such as blindness, nerve damage (neuropathy) and kidney damage.

DIABETES TREATMENT: Using Insulin

Diabetes is treated by: regular injections of insulin. These are given daily or several times a day, depending on the type of insulin used.

DISCOVERY OF INSULIN
In 1921 the Canadian scientist Friedrik G. Banting and Charles H. Best discovered insulin which lowers blood sugar. They extracted insulin from animal pancreas.(animal insulin)

DISCOVERY OF HUMAN INSULIN (HUMULIN)

In 1978, scientists from the biotechnology corporation Genentech in San Fransisco, using a genetically manipulated plasmid of E. coli bacteria, suceeded in producing insulin with the same amino sequence as in humans (human insulin).Recombinant DNA technology is used when producing human insulin.

The Genetics of Recombinant DNA

Recombinant DNA is the process of merging genes from one organism with the genes of another organism. This process involves using restriction enzymes to isolate a gene and then place

it on a vector plasmid. Then it is inserted

into a host cell such as bacteria. The altered plasmid taken up and incorporated into the genome. When the bacterial cells

divide they pass on the new information to

the next generation. This produces clones of bacteria that produce large quantities of the new protein.

Insulin Production

Cut isolated DNA from human cellls with a restriction enzyme

6 Join together plasmid and human fragment

 Then, the recombinant plasmid is mixed with bacteria.

One cell with the recombinant plasmid

8th step when gene cloning takes place. The single recombinant plasmid replicates within a cell.

Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid and the human insulin gene.

Route to the Production by Bacteria of Human Insulin

The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant human insulin gene.

Scale Up Step
In the process of comercial production of proteins, cell cultures must be scaled up to produce enough protein to meet demand. Since only a limited number of cells can be grown in small petri dishes or flasks, the cell culture can be transferred to large containers called bioreactors, which are involved in the manufacturing process.

Raw Materials and Required Instruments in the Synthesis of Human Insulin

Escherichia coli is the most widely used type of bacterium, but yeast is also used. Researchers need the human protein that produces insulin. Manufacturers get this through an amino-acid sequencing machine that synthesizes the DNA. Also necessary to synthesize insulin are large tanks to grow the bacteria, and nutrients are needed for the bacteria to grow. Several instruments are necessary to separate and purify the DNA such as a centrifuge, along with various chromatography and x-ray crystallography instruments.

VACCINES
Vaccines stimulate the immune system and provide protection against particular diseases.

The first vaccines were made with inactivated

(killed) or weakened virus unable to reproduce

in the body but sufficient to provide immunity

upon future exposure to the live virus.

DNA VACCINES
Vaccines are also created with recombinant proteins. Scientists use genetic engineering to create recombinant vaccines DNA vaccines are a new form of gene therapy wherein a few genes are extracted from a virus and injected into people for curing many forms of disease and illness. Unlike the typical process which may take up to six months, DNA vaccines can be ready in less than three months. DNA vaccines is used cancer treatment and in future AIDS treatment.

DIAGNOSTICS (tehis,tan)
Scientists use recombinant DNA technology to produce a number of diagnostic tests for diseases, including tests for cancer, hepatitis and AIDS. One type of gene polymorphism, Single Nucleotide Polymorphisms (SNPs), can be used as a diagnostic tool.

SNPs (Single Nucleotide Polymorphisms)

Normal DNA

Polimorfizm

Mutant DNA

Functional protein

Functional protein

Nonfunctional Protein or Loss of Protein

SNPs can be used to build a profile of a persons tendency to various diseases. (cancer, heat attack risks etc.)

SNPs using in early diagnosis

PHARMACOGENETICS
Pharmacogenetics is a relatively new field of study which deals with how genetic factors influence the way individuals respond to drugs. It is hoped that this type of study will help scientists optimize drugs, and perhaps even one day create entirely individualized treatment programs based on individual genetics.

Pharmacogenetic
Pharmacogenomics is a key term, referring to the use of information about the genome to develop drugs in pharmacogenetics. Pharmacogenomics is being used for all critical illnesses like cancer, cardio vascular disorders, HIV, tuberculosis, asthma and diabetes.

 Pharmacogenomics is aimed to predict if a patient will have a good response to a drug, a bad response to a drug, or no response at all.

In Future

Personalized Medicine Pharmacogenetics is an important branch in the future for biomedical developments. It provides special treatment for each patient not average patient profile. Pharmacogenomics and gene therapy will prevent the cancer people before it develops. Stem Cell theory is integrate into the diseased tissue, replace diseased cells and reverse the effects of the disease. Nanomedicine is used nanoshells to selectively target and destroy cancer cells at the cellular level.

20 New Biotechnological Breakthroughs that will Change Medicine

1.Decay-Fighting Microbes (rmeyen diler) 2. Artificial Lymph Nodes(yapay lenf dmleri) 3. Asthma Sensor (nano astm sensr) 4. Cancer Spit Test (tkrkle kanser testi) 5. Biological Pacemaker (biyolojik kalp pili) 6. Prosthetic Feedback (hissedebilinen protezler) 7. Smart Contact Lens (akll kontak lensler) 8. Speech Restorer (yeniden konuturan ara) 9. Absorbable Heart Stent (eriyen damar stenti) 10. Muscle Stimulator (elektrikli kas uyarc)

11. Nerve Regenerator (yeni sinir ular) 12. Stabilizing Insoles (dengeleyici ayakkab) 13. Smart Pill (akll ila) 14. Autonomous Wheelchair (akll tekerlekli sandalye) 15. Gastrointestinal Liner (zayflatan mide astar) 16. Liver Scanner (5 dk da karacier testi) 17. Nanoscale Adhesive (tamir eden nano yaptc) 18. Portable Dialysis (tanabilir yapay bbrek) 19. Walking Simulator (yrme simlatr) 20. Rocket-Powered Arm (roket motorlu kol)

Pancreatic Tooth for Diabetes

Control of sugar in blood is an important factor in diabetic end-organ damage.

Artificial pancreases provides responsive glycaemic control that can reduce

enormous personal suffering and socio-economic costs of diabetes. However, they share the defects of limited operational life, due to depletion of stores or failure of component parts.

A pancreatic tooth design provides

accessibility

for

the

purposes

of

replacement. In addition, the mouth also provides a sheltered location, is more resistant to diabetic changes and less tendency changes (deri to than thermoregulatory subcutaneous

altndaki)

tissues, and is adapted to cope with the occasional pathogen load.

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