LSSI July 2010matsc.org/Unprotected Docs/Teacher PPTs/10 LSSI ppt.pdfmost successful recombinant protein companies in the world. • Concentrating in oncology, neurology and immunlogy

LSSI July 2010

• The LSSI program is currently being funded through San Diego Workforce Partnership by industry and education partners, specifically the Amgen Foundation, Biogen Idec Foundation, Genentech and Southern CA Biotechnology Center (SCBC) at Miramar College.

Externships

• Biogen was founded in 1978 by a group of noted biologist.

• Biogen and Idec merged in 2003 to create one of the most successful recombinant protein companies in the world.

• Concentrating in oncology, neurology and immunlogy it has become a 4 Billion Fortune 500 company.

• Biogen Idec, a great supporter of science education, sponsors the Community Lab here in San Diego and also has a training center for teachers K-12 in its North Carolina facility.

Current Therapies• Tysabri therapy for Multiple

Sclerosis• Avonex therapy for Multiple Sclerosis• Rituxen therapy for non Hodgkin’s Lymphoma

and Rheumatoid ArthritisSanDiego is a R&D facility for oncology. They take the protein from discovery to small scale production protocols.

• The use of column for purification, we did the same thing only using a smaller column.

• Growing cells in a shaker of liquid medium, ours was a 250mL flask, again theirs was a little larger.

• Cancer resistance in CHO cells (I now know what that means), and we got to see the change in their morphology.

• Flow cytometry…sorting and placing one cell in over 300 wells!!!!!!!.

• And scale up, all that and MORE in one building. AND DON’T FORGET THE CAFÉ!

• Established in 1998• A pioneer in the use of high throughput receptor - based

assays to discover new flavor ingredients• FEMA GRAS* approval received on the following

products:• 4 savory ingredients (used by Nestle and other

partners)• Sucralose enhancer (used by Firmenich)• Sucrose enhancer (used by Firmenich)

*FEMA GRAS: Flavor and Extract Manufacturers Association; Generally Recognized As Safe

Active Areas of Research:• Determining the exact mechanisms of salt receptors

in order to develop salt enhancers allowing the reduction of salt in foods

• Discovering new cooling ingredients

Active Areas of Development:• Fructose enhancers undergoing optimization• New sucrose enhancers currently being taste tested• Preparing to submit bitter blockers for approval by

FEMA

Through the use of enhancers, the amount of sucrose can be reduced by up to 50% and the amount of sucralose can be reduced by up to 70% in certain food/beverage products. Wow!

Senomyx is partnered with top companies in the food and beverage industry including Nestle, Campbell, Firmenich, and possibly PepsiCo in the near future.

We saw a direct classroom connection. In the labs we observed the use of high through-put assays. LSSI has equipped us to run an ELISA (enzyme-linked immunosorbent assay) within our classrooms.

• Gen-Probe is An Innovative Growth Company in Molecular Diagnostics that has been providing appropriate molecular solutions

for DNA testing to laboratories since 1985 has a proven track record in providing technically

superior products of the highest quality to test for various infectious diseases

has an 11% to 12% share in the Molecular Diagnostics Industry

• Gen-Probe was founded in 1983 & Acquired by Chugai Pharmaceutical Co. Ltd. For $110 million in 1989. They received ISO certification in 2006 and were awarded The National Medal of Technology and Innovation bestowed by President George W. Bushin 2004.

Nucleic Acid Tests for the detection of Prostate Cancer

Expanding Women's Health Franchise for detection of high-risk strains of HPV and Trichomonas vaginalis.

Instrumentation: TIGRIS a high volume automated molecular diagnostics instrument and the Panther Platform an extension of Gen-Probe's TIGRIS® instrument

Human Disease Diagnosis: STDs, HIV, Hepatitis,West Nile Virus, Mycobacteria, Group A Streptococci, Fungi, Oncology.

• Established in the 1960s by Jonas Salk, M.D., the developer of the polio vaccine.

• His goal was to establish an institute that would explore questions about the basic principles of life and the future of humanity.

• In December 1959, Salk and architect Louis Kahn began a unique partnership to "create a facility worthy of a visit by Picasso."

• In March 1960, Salk and Kahn were given the Torrey Pines land.

• 5 of the scientists trained here have won the Nobel Prizes. 3 are currently there.

• Bioinformatics – Razavi Newman Center• Center for Cytometry and Molecular imaging – CCMI• Functional Genomics• Peptide synthesis – for Institute Scientists• Proteomics – protein ID by mass spectrometry• Stem Cell Core – training & physical space• Transgenic – Mice, IVF, & cryopreservation• Viral Vector – design, consult, & production for vector

systems

• CIRM awards Salk scientist $15.6 million to

• Slow or halt motor neuron degeneration,

• Applications in ALS• The amazing sculptures by Dale

Chihuly • The staff photo contest• The animal lab • Thank you David Gonzales• The student interns• The beating stem cell• Thank you Travis Berggren

• 1st Biotech; Founded in 1976; Worldwide Industry• Second Largest Biotechnology Company

• Research, Development and Production• 1,100 researchers• 17 major drugs approved from 1978 to present• 35% of world manufacturing capacity for protein drugs

• Whole company-3 focus areas• Oncology• Immunology• Tissue Growth and Development

• Oceanside Manufacturing Facility-Purchased in 2005

• Produces large quantities of Avastin-inhibitsangiogenesis which slows the growthof carcinomas in colon and rectumtumors

• Will be switching to manufacturing Actemrafor Rheumatoid Arthritis.

• More opportunities for various education levels at a manufacturing site

• Licensed to produce: Rituxin, Raptiva, Actemera, and Avastin, but only produceone at a time

• Scale! Have several 10,000 liter tanks• Green:

1. Decrease electricity by 15% by 20142. Initiatives such as Green Genes to research issues and educate employees3. “Gravity Manufacturing”

Company Information & History:• Invitrogen founded in 1987 and

acquired Applied Biosystems to become Life Technologies

• Global biotechnology tools company dedicated to improving the human condition through research, development, & production

• Provide premier systems, consumables, and services that save time, money, labor, and reduce non-green & biohazard waste

• 3,900 patents and exclusive licenses

Products:• Antibodies & Secondary Detection Reagents • Beads & Microspheres • Buffers & Chemicals • Cell Culture & Transfection Reagents • Columns, Resins & Plates • Competent Cells & Strains • Controls, Standards & Ladders • DNA Vectors, Clones & Libraries • Eukaryotic Cells • Gels, Fractionation Strips, & Membranes • Instruments, Software, & Accessories • Kits & Assays • Labeling & Detection Products • Literature • Primers, Primer/Probe Sets, Oligos &

Nucleotides • Proteins, Enzymes, & Peptides • Slide & Chip Arrays

Services:• Molecular Biology Services • Antibody Production, Assay Development &

Labeling Services • Screening and Profiling • ADME/Tox Services • PD-Direct® Bioprocess Services • Custom Media Production (cGMP) • GIBCO® MediaExpress™ (non-cGMP) • Cell Culture Services™ • Licenses, Terms, and Conditions

Research and Development:• Assay & Antibody Development Services• Screening and Profiling• ADME Development Services• PD-Direct® Bioprocess Services• Custom Media Production (cGMP)• GIBCO® MediaExpress™ (non-cGMP)• Molecular Biology Services

• Awesome E-Gel® Precast Agarose Gel Cartridges• Mother Bases, & Transilluminators for Electrophoresis

• Fast, easy, and safe!!!

• #1 Pharmaceutical Company in the world• $50,000,000 gross in 2009• 125 medications• Founded in 1849

• Charles Pfizer• Charles Erhart

• “Learning that in R&D, accepting to not succeed 99.9% of the time, and hoping for that .1% because that could mean saving the lives of 1 or a million makes it all worth it.” – Tessa

• “It was cool to see the concepts I teach in school being applied in industry.” - Tim

• “Wow, using safer methods and chemicals, less waste or none.” – Roger

• “My son had lymphoma with a stem cell transplant. Pfizer will help my uninsurable son get access to Pfizer medicines that could mean the difference between life and death.” – Laurie

• Began as Mycogen• DOW acquires Mycogen for

Pseudonomas• Pfenex spins off from DOW

December, 2009

• Pfenex is involved in functional recombinant protein production and expression using psuedonomas bacterium. P. fluorescens is a robust fermentation organism delivering high cell densities and product yields.

• Dow acquired the psuedonomas platform from Mycogen Corp in 1998 and Pfenex was formed in 2009.

• Pfenex manufactures therapeutics such as G-CSF, IFN and hGH and vaccines (undisclosed). The proteins they manufacture are of 4 types: Reagent proteins, Research proteins, Lead proteins, Biosimilars.

• Seeing robotics in action was amazing• Go WALL-E

• The employees lived their jobs, they loved science• Smarter not harder• CEO mtDNA/CSI Presentation

• Conatus was founded in 2005 by Idun management when the company was purchased by Pfizer,relocated, and did not re-hire employees. It is a “virtual” biotech research company (acquires partially developed drugs, runs trials, and sells the drug).

• Conatus researched potential drugs for purchase and found a discarded project for rhumatoid arthritis showed promise as hepatitis C (cause of liver disease) treatment.

• Conatus focuses on damage caused by hepatitis C virus.• Activated stellate cells release proteins to repair

damaged liver cells.• Hep. C virus activates stellate cells in undamaged

liver cells. • Causes inflammation, scar tissue and protein

buildup.• Conatus developed CTS-1027 (2007)

• Inhibits MMPs (matrix metalloproteases) that cause inflammation.

• Most recent trial includes a combination of three antiviral drugs as a therapy.

• Very small company• Everybody knows everybody

• A virtual science company• Very adventurous employees• Reusing shelved products to try find new uses• Finding treatment for Hepatitis C by combining three

drugs

THANK YOU!

VERY MUCH!

AmgenBruce-Wallace

Biotechnology Lab Series

Ever wonder what really goes on in a Biotech lab?

This series is for you!

Lab 2: Restriction Treatment of the plasmids pARA and pKAN-R

They use special proteins called restriction enzymes to cut genes into pieces.

It starts with…. Lab 1: Introduction to Pipetting and Gel Electrophoresis

LOOK! Teachers are actually pipetting! They are introduced to pipetting techniques and learn how to run a gel

We use these micropipeters to measure the very, very small

amounts!

Ever wonder what scientists are really doing in a lab?

How DO scientists isolate a gene?

Lab 3: Ligation of pARA/pKAN-R Restriction Fragments

We took the fragments we cut from the two plasmids and made a new one called a recombinant plasmid. This new plasmid should have a fluorescent protein AND be ampicillin resistant.

LOOK:Teacher cutting without scissors

Do you like to cut and paste?

Lab 4: Confirmation of Restriction and Ligation

We looked at bands on an agarose gel to determine if our plasmid was present and the correct size. It will look something like this!

How can you visualize something you can’t see?

They need agitation! SHAKE, SHAKE, SHAKE!

Lab 5: Transformation of E.coli with a recombinant plasmid

We used different media to encourage E. coli to take in the new plasmid. The presence of pink colonies confirmed that the bacteria made the new protein from the plasmid.

Lab 6: Preparing an overnight culture of E.coli

We grew bacteria overnight by agitating it with nutrient-rich media in a flask.

YES! It worked….

glowing red colonies!

Did you know that bacteria need food too?

How is foreign DNA put into a bacteria?

Mix of all the proteins …..

… then….

…You have purified red fluorescent protein!

… it is PINK!

Lab 7: Purification of mFP from the overnight culture

We exploded the bacteria cells and used a column full of resin to separate the components. We were able to locate

and isolate the product; a red fluorescent protein.

How do scientist actually get the product out of the bacteria?

Lab 8: Genomic DNA Extraction and Amplification through PCR

WOW! That is your actual DNA

Using a PCR machine, we made multiple copies of our DNA. We then used gel electrophoresis to determine our genotype based on how the fragments are separated in the gel.

PCR is one of the most powerful techniques used in molecular biology.

Ever wonder how DNA fingerprint works?

Pos. Control Neg. Control Positive!!!! Maybe???

ELISA (Enzyme-linked Immunosorbent Assay)

A pregnancy test uses antibody interactions and specific chemicals to create a color change when the pregnancy

hormone is present.How does a pregnancy test really work?

Personal Experiences

We are...

We are students…

We areDNA…

We aredialed-in…

We are balanced…

We aregellin’…

We are on overload…

We areAMAZED…

We arepositive +…

We arethirsty…

We arehonored…

We aregreen…

We areprecise…

We are QC…

We are phat…

We are inquisitive???

We are complementary…

We aretwisted…

We arevarious molecular

weights…

We are grateful!!!THANK YOU!!!

We are the teachers of future biotechnologists!