The LSSI program is currently being

funded through San Diego Workforce

Partnership by industry and education

partners, specifically the Amgen

Foundation and Southern CA

Biotechnology Center (SCBC) at

Miramar College.

During our 12 day LSSI experience, we

visited 8 different biotechnology

companies in San Diego. These

companies include Pfizer, Pharmatek,

Sapphire Energy, White Labs,

Senomyx, Salk, Pfenex and Conatus.

The following slides are an overview of

what we got to see and what we learned!

Largest research based

pharmaceutical company

Leading portfolio

of products and

medicines that

support wellness

and prevention

Our Visit

Welcomed with a wonderful lunch!

Our host gave us an overview of the pharma-industry, an overview of drug discovery and how drugs reach the market place.

Following the discussion, we toured the labs where we saw equipment

and techniques applied in

drug discovery and

research.

Connection to Amgen Labs

At Pfizer we observed the use of PCR, gel electrophoresis, and protein synthesis using insect cells to produce a recombinant protein.

This connected to Amgen Lab in creating a recombinant plasmid, transformation of pARA-R into E. coli and purification of mFP

Compound

Evaluation

Analytical Methods

Development

Formulation

Development

GMP Manufacturing

Stability Testing and

Storage

Cytotoxic and High-

Potent Development

http://www.pharmatek.com/index.html

Amgen Lab 4-Confirmation of

Restriction and Ligation

◦ Pharmatek uses HPLC to confirm the products

they make contain the correct amounts of the

various components

Extension of the Amgen Lab Series

◦ It demonstrates the importance of formulation

and delivery to the body.

• Algae-based Green Crude oil production

• Upgrade algae biomass into crude oil

• Finding and manufacturing a renewable energy

source that benefits the environment

• Drop-in replacement fuels for gasoline, diesel, and

jet fuel

• A company with a team of investors

entrepreneurs, scientists, & concerned citizens

Renewable

Low carbon footprint

America’s energy independence

Price-competitive

Fits into our existing energy infrastructure

The Benefits of Algae-Based

Green-Crude Oil Production

Connections to Amgen Labs Growing algae in different situations to

test each strain’s viability

Polymerase Chain Reaction – to screen

for the strains of algae and their

concentrations and size

Research & Development in harvesting

cells and analyzing the algae (determining

the DNA sequence)

White Labs Inc. was founded in 1995 by Chris and Lisa White, who had experience growing yeast in labs.

Make beers, supply breweries, wineries and distilleries with fresh yeast and related products, including analytical services

White Labs: Products & Services

Production of yeast process:

strain selection plate

working plate liquid cultures*

yeast propagator (big scale)*

500 different flavor compounds

Analytical services: alcohol

content, calories, flavors, QC.

Partners with non profit water

treatment. Produces cleaner

water than most water

treatments.

* Lab tested

Connections to

Amgen Labs

Using strain selection techniques: DNA profiling, PCR, growth of the best strain on plate then in liquid media, sterile techniques.

Class tours are welcome!

Applying sensory science to create

artificial flavors and flavor enhancers.

• Founded in 1998

• Founders:

• Lubert Stryer—neurobiologist, Stanford Univ.

• Charles Zuker—neurobiologist , Columbia Univ.

• Roger Tsien—molecular engineer, UCSD, Nobel

Laureate, 2008.

Products: • Savory taste additive reducing

or replacing MSG

• Sweet taste modifiers reducing

need for sucrose, sucralose, &

high fructose corn syrup

• Bitter blockers

• Cooling taste product

Research: • Identify salt taste receptor

• Discover new taste enhancers

• Senomyx protocol includes assaying for potential taste

enhancers, similar to the ELISA (Enzyme-Linked Immuno

Sorbant Assay) lab

• Identified flavor enhancing substances are purified using

column chromatography techniques, similar to Amgen lab #7

protocol.

Salk institute labs

started by Jonas Salk

(discoverer of polio

vaccine) in 1960.

350 graduate and

postdoctoral scholars;

1200 staff from 36

different countries.

Inside Salk

Research Domains at Salk

Genetic medicine, dynamic

brain, healthy aging, plant

biology, Waitt Advanced

Biophotonics center, Cancer.

Dr. Geoff Wahl, Gene

Expression Laboratory;

identification of cells

initiating cancer; last

discovery on the genes in

breast cancer cells, potential

targets for cures.

Animal Research

Salk Animal Facility (mice, guinea pigs,

rabbits), Cancer Research Animal Facility,

Non Human Primates. Managed by

Brandon Becker.

CONNECTIONS TO AMGEN LABS

PCR, Gel Electrophoresis, Protein Transformation, DNA Amplification!

Salk Mobile Science lab for middle schools.

March of Dimes High School Science day initiated 30 years ago by Jonas Salk.

Life Science Summer Institute.

A biotechnology company committed to developing

biosimilars and innovative vaccines in order to address unmet

and growing global healthcare needs.

Utilizing Pseudomonas fluorescens, rapid

automated screening, and recombinant DNA

techniques, Phenex produces biosimilar

therapeutics, vaccines, and reagent proteins

rapidly, in high yield, and that are of high quality,

activity, and solubility. They also provide strain

engineering services for other companies.

Biosimilars:

• Ranibizumab

• Certolizumab pegol

• Interferon beta-1b

• Human Growth Hormone (hGH)

• Granulocyte Colony-Stimulating Factor (GCSF)

Vaccines:

• Recombinant Protective Antigen Anthrax (rPA) Vaccine

• Malaria Vaccine

Cut and ligate a desired plasmid gene and use it to transform

P. fluorescens.

Maximize protein production via assaying P. fluorescens strain

protein yields and customizing optimal growth conditions

(e.g., pH & temp.).

Test for product via electrophoresis, HPLC, and the mass

spectrometer.

Purify protein product via filtration and chromatography.

This general mode of operation closely resembles our

entire Amgen lab series, which also involved generating

& verifying recombinant DNA, transformation,

proliferation, protein harvest, & analysis.

Conatus is focused on the development of therapeutics for liver disease and cancer.

In 2010, Conatus acquired Pfizer's Idun subsidiary, which brought Emricasan, a Phase 2 drug candidate for fibrotic liver disease; IDN-13389, a preclinical candidate for oncology indications; an agreement with Abbott for an oncology drug in Phase 2 clinical trials; and IP in the field of apoptosis.

They are now trading on NASDAQ

Ticker: CNAT 9.50 (- 1.50)

Conatus

introduced us to

all the soft skills

that are important

to all biotech

companies.

They showed us

how a small

biotech company

operates!

Lab 1: Microvolumetrics and Pipetting

Goals:

◦ To familiarize students with small

volume solutions

◦ To practice using a micropipette

◦ To practice gel electrophoresis

Lab 2:

Restriction Analysis of pARA and PKAN-R Goals:

◦ To examine the role of plasmids in molecular biology

◦ To examine the role of restriction endonucleases (restriction enzymes) in molecular biology

◦ To initiate the first step in producing a recombinant DNA molecule

Lab 3: Ligation of pARA and pKAN-R

Restriction Fragments Producing a

Recombinant Plasmid, pARA-R

Goals:

◦ To teach students how DNA from separate

plasmids can combine to produce novel plasmids

◦ To isolate the recombinant plasmid pARA-R for

future lab use.

Lab 4: Confirmation of Restriction &

Ligation Using Agarose-Gel

Electrophoresis Goal:

◦ To confirm that the restriction enzymes digested

pKAN-R and pARA and that a new plasmid pARA-R

was produced by comparing digested plasmid, and

undigested plasmid to ligated plasmid.

Lab 5: Transformation of E. coli with a

Recombinant Plasmid Goal:

◦ To make cells that are genetic factories for a

recombinant protein.

Lab 6:

Production of

Recombinant

Protein

Goal:

◦ Use prokaryotic cells as factories for a

recombinant eukaryotic protein.

Lab 7: Purification of mFP

Goal: to prepare the cell lysate from the

overnight liquid culture and purify the

mutant fluorescent protein.

Lab 8: Amplification of the tPA

Locus using the Polymerase Chain

Reaction (PCR) Using a very

powerful technique to amplify buccal cell DNA and determine student genotypes.

The PCR technique was invented by Dr. Kary Mullis in 1983. He was awarded the Nobel Prize in Chemistry in 1993.

ELISA

Enzyme-Linked ImmunoSorbent Assay

Detecting the presence of antibodies or antigens

Uses the binding of antigens and antibodies to see a change, such as color

Tests for Pregnancy, HIV, West Nile, Hepatitis,

Protein recognition

DNA Extraction from Wheat Germ

Let us tell and show you what our favorite

experiences were, during LSSI this summer!

BEST ASPECT of LSSI..

Reuniting former students and teachers!

I had Mr. Kepper as my

10th grade in 2001! I still

can’t call him by his first

name. Biology teacher

Tina Tom School: Chula Vista

Middle School

Teaches: General Science 1/

Health 1

Years Teaching: 3

Favorite Scientist: Ivan Pavlov

Chris Osborne School: Mt San Jacinto College

Teaches: Biology

Years Teaching: 27

Favorite Scientist: John Sanford

Christine Arrabrit School: North County Trade

Tech High School

Teaches: Earth Science,

Biology, Chemistry, and

Physics

Years Teaching: 1

Favorite Scientist: Richard

Morimoto

Josh White

School: Vista Murrieta High

School

Teachers: Biology (AP), Marine

biology, Anatomy and

Physiology

Years teaching: 8

Nicole Clymer School: Escondido High School

Teaches: Biology, Oceanography

Years Teaching: 3

Favorite Scientist: Rosalind Franklin

Cesar Lopez School: Indo High School

Teaches: AP Biology and

Chemistry, Vet Science, Small

Animal Care, Horticulture

Science

Years Teaching: 2

Favorite Scientist: George

Washington Carver

Jessica Ullyott School: Gompers Preparatory Academy

Teaches: Biology

Years teaching: 3

Favorite Scientist: Don Herber

(Mr. Wizard)

Katie Martin School: San Diego High-SciTech

Teaches: Biology, Chemistry,

Human Body Systems

Years Teaching: 1

Favorite Scientist: Gilbert Lewis

Jeff Kepper School: Helix Charter High School

Teaches: Biology and Oceanography

Years Teaching: 21

Favorite Scientist: Robert Ballard

Bekah Walker School: Rock Academy

Teaches: Biology, AP Biology, Life

Science, Anatomy and Physiology

Years teaching: 5

Katie Johnson School: Mt San Jacinto College

Teaches: Biology and Humans and

Scientific Inquiry

Years teaching: ½

Favorite Scientist: Alfred Russell

Wallace and Charles Darwin

Alec Barron School: Del Lago Academy

Teaches: Biochemistry and Engineering

Years teaching: 10

Favorite Scientist: Erwin Schrodinger

Gina Saclauso-Caro School: Chula Vista Middle School

Teaches: Life Science/Health

Years Teaching: 30

Favorite Scientist: Madame Currie

Beth Mueller School:

Foothills Christian High

School

Teaches:

Chemistry, Biology, Anatomy

Years teaching: 5

Favorite Scientist:

Gregor Mendel

Alicia Johal School: Mar Vista Academy

Years: 1st year teacher!

Teaching: 8th Grade General Science

Favorite Scientist: Charles Darwin

Thank you Sandy!

Thank you for your presentations, insight, and

knowledge!