Download View Powerpoint Presentation - Northeast Biomanufacturing Center

How to Start a Biotechnology
Program
Mary Jane Kurtz, Ph.D NBC2 Consultant
[email protected]
Why Biotechnology?
A number of good reasons
1.
2.
3.
4.
Academics
Technical skills
Biotechnology as a future career
Biotechnology is exciting
Biology can be seen with a molecular perspective
Can answer questions about ourselves
Can provide new medical aid to the afflicted
Getting Support from Your
Administration
This is important to the future success of your
new program
Allows for obtaining outreach to new students,
space to grow, and expenses for needed
equipment
Supports the teachers who are involved in
program
Academic Advantages
 Allows students to do up-to-date hands-on
experiments
 Demonstrates the integration of all sciences,
math, and engineering through experiences in
the laboratory
 It develops writing skills for clarity, focus and
documentation
Technical Advantages
 Curriculum can introduce career tracks for
students in a developing scientific area
 Using hands-on activities, students have a
opportunity to develop skills needed in the
workplace at all levels of formal training
 Potential career paths flowing from the
knowledge gained is broad based
Biomanufacturing Curriculum:
National Standard Concepts Covered
• Math, Biology,
Chemistry in Curriculum
–
–
–
–
Measurement
Solutions
Enzyme reactions
Transformation of cells
with DNA
– Forces used in
centrifugation,
electrophoresis etc.
• National Academy of
Sciences Standards
– Unit of Math & Science
– Structure/properties of
matter
– Chemical
reactions/conservation
of matter
– Cell structure and
functions, heredity
– Motions and forces
Biotechnology
A plus for Teachers and Students
 Integrated science education: Science Technology
Engineering and Math (STEM)
 Career pathways towards work/school focused on
science with thousands of new jobs predicted in the
next few years
 Laboratory based activities = increased interest
– More exciting ways of introducing concepts
– Hands-on learning is more inclusive
– State-of-the-art laboratories
Biomanufacturing vs Biotechnology





Offers diverse career pathways
Hands-on learning of science
Understanding of good laboratory practices
Stability in career pathways
Assistance in obtaining college degrees
through company support
 Both highly academic and technically oriented
students
Ten Technician Jobs Anchor
Ten Biomanufacturing Departments
• Facilities/Metrology
• Validation
• Environmental Health and
Safety (EH&S)
• QA
• Upstream Processing
• Downstream Processing
• QC Microbiology
• QC Biochemistry
• Process Development
RESEARCH &
DEVELOPMENT(pre-clinical):
Discovery Research
OPERATIONS:
QUALITY:
Process development,
Manufacturing
& Production
Quality Control
& Assurance
Discovery Research
Process
Development
Director Supervisor &
Process Development
Technician
Quality Control
(QC)
Bioinformatics
Scientist
Engineer
Analyst
Programmer
Manufacturing
& Production
Supervisor
Associate
Technician (Operator)
Instrumentation Tech
Calibration Technician
Facilities Management
Manager
Facilities Technician
Shipper/receiver
Chemistry
Chemistry QC
Analyst
QC technician
Microbiology
Microbiology
QC analyst
QC Technician
Quality Assurance
(QA)
Documentation
Specialist
QA Documentation
Coordinator
Clinical Research
Clinical Research Manager
Regulatory Affairs
Manager
Associate
Data Manager
Business Development
Director of Business
Development
Administration
Human resources
Safety Manager
•Careers in red indicate
entry level positions
All above require 2-4 year
Entry level positions require
an Assoc deg or certificate
Higher entry levels require a
BS, MS, PhD or Engineering
degrees
Can require medical, nursing
or business degrees
Senior Scientist
Scientist III,II,I
Research Associate
CLINICAL RESEARCH:
Clinical Research:
Regulatory
Affairs
Clinical Research
Clinical Research Manager
Student Success Stories
Students completing courses with certificate or
Associate degrees in Biomanufacturing:
www.biomanufacturing.org
http://www.biomanufacturing.org/
1. How to Start A Biotechnology
Program
(After getting support from the administration)
Gain the support the Industry in your Area:
–
–
–
Find out the location and focus of each of the biotechnology/support
companies within (10) miles of your school
Understand the desired skills individuals (such as your students)
should have (Most important of these are soft skills)
Organize an advisory group that can assist you
in curriculum building towards skills needed
This will be a source of information as well as help with technical
supplies
2. Find or Develop a Curriculum
Resources
High School Level-Entry Level Community College:
Protein is Cash:
Northeast Biotechnology Center and Collaborative , NBCC:
www.biomanonline.org, Biomanufacturing.org
Biotechnology: Science for the New Millennium:
Ellyn Dougherty, EMC publisher, www.emcschool.com
Basic Laboratory Methods in Biotechnology:Textbook and
Laboratory Reference:
Lisa Seidman et al, Prentice Hall, ISBN 013-795535-9
Align Curriculum with
Program Goals
 What are your goals?
– Two year associate degrees transfer to four year
college
– Two year associate degree and work
– Certificate and immediate work
 Research and Development or
Manufacturing?
 Goals can vary but should allow majority
of your students to participate in.
Four year college career pathway
Biology Undergraduate
Other Undergraduate
or Engineer
Biomedical Ph.D student
Other Ph.D student
Junior Postdoc &
Senior Postdoc
Industry
Other Science Position
Staff Scientist
Assistant Professor
Independent Investigator
Intro to Biotechnology for High School/CC
1
2
3
4
5
6
7
8
9
10
11
12
Introduction to Biotechnology
Use of bacteria, enzymes and
fermentation by yeast
Metrology
Validation of Pipets and Calibration
SOP ‘s &
Documentation of Testing
Basic Microbiology
Basic Aseptic Technique and Testing of
Unit 1
Bioload in Biomanufacturing Areas
Gowning up
Basic Microbiology
Identify of Microbe
Identification of Bacterial Types
Morphology
Unit 2
and Electrophoresis
Transformation of Cells
PGlO transformation of
With DNA
Bacteria
Spectrophotometry
Protein determination of samples
By Bradford and UV analysis
Genetic Transformation of Bacteria
E. coli transformation with pGLO
plasmid
The Rain Forest:How do we find new
Growth of pGlo transformed bacteria
pharmaceuticals ?
and isolation of the p-GLO product
Proteins and their purification
Isolation of casein from milk
Using solubility and filtration
techniques
Good Manufacturing Practices
GMP activity with popcorn
Documentation
And use of Documentation
Chromatographic separation and how it Use of ion exchange and hydrophobic
works
columns in
Isolating GFP
QC Sampling of Product to
Polyacrylamide Gel Electrophoresis
Determine its
Purity
Biotechnology Curriculum
2. Associate Degree or Certificate Program
Resources:
Introduction to Biomanufacturing: Global
Biomanufacturing Lab manual
NBC2, at www.biomanufacturing.org
Introduction to Biomanufacturing: Textbook,
NBC2, at www.biomanufacturing.org
Introduction to Biotechnology Lab Manual
ATE central: Linnea Fletcher, et al ISBN BITC1311006
Techniques in Biotechnology
Week
Lecture and Topic
Laboratory
for Community
College
1
Acids, Bases and
Buffers
Protein &
Spectrophotometry
Preparation of
Buffers
Determination of
Protein in solution
2
Enzyme Kinetics
Kinetics of LDH
Protein structure
Km of enzymes
3
Purification by pH
Column
Chromatography
Precipitation of pro
tein
and Affinity
chromatography
4
Enzyme Purification Purification of LDH
by Ion Exchange
using a IEX column
5
Identification of
Proteins
Concentration and
Electrophoretic
Techniques
6
High Pressure
Chromatography
Quality control
Case study of Two Different
Programs
College Degree
• Requirements will be more
demanding in math, science
• Comprehension, graphing
&computer work is good
can work independently
• Has appropriate soft skills
and can work well as a team
member
• Is good at trouble shooting
problems in labs
High School or Certificate
• Entry level math is prealgebra or algebra 1
• Science background is
sketchy (lacks chemistry)
• Desire to be placed into
a position after graduation
• Likes science but needs
reassurance
• Needs to have SOP to carry
out work
High School Curriculum
A Workshop to Instruct Teachers and Students
in
Biomanufacturing and the Bioeconomy
Career Tracks: Protein is Cash
Introduces
 Upstream Processing
– Production of pGLO into protein by transformed cell
 Downstream Processing
– Separation of cellular debris and cell supernatant
– Purification of pGLO protein by Chromatography
 Quality Control
– Identification of protein product by electrophoresis
 Discovery Research
– How new drugs are discovered
 Biofuels
1.
2.
3.
4.
5.
6.
Metrology
Transformation of Bacteria
Upstream Processing
Downstream Processing
Quantitative Analysis
Discovery Research
$Protein
is Cash$ Local Teachers
Workshops
Biomanonline.org
Day 1 : Metrology/Instrumentation
Activities
Outcomes
• Calibration of top balance
• Introduction of Good Manufacturing Practices
• Verification of
pipet performance
• Pipetman Challenge
• SOP and documentation
• Confidence in
measurements by instrumentation
Metrology
Day 2 Transformation of Bacteria
Activities
• Addition of pGLO plasmid to
bacteria in Ca++ solution
• Heat /shock the mixture
• Plate cells onto selective
Luria broth agar + ampicillin
• Incubate overnight at 37oC
• Note: arabinose acts to turn
on the gene which starts
production of pGLO protein
Outcomes
• Selection of cells by growing
on ampicillin plates
• Only transformed with cells
will survive due to amp-r
gene
• Selected colonies will
multiply in Luria broth at
37oC overnight
• Aseptic technique and
proper disposal of bacteria
Day 2 E.coli Transformation
with pGLO plasmid
Ampicillin resistance gene (Ampr)
and target gene on bacterial plasmid
Cell division
Bacterial clones
Only E. coli containing plasmid
survive on Ampicillin plates
Transformation mixture is plated
on to agar plate containing
Ampicillin
Results of Inserting Foreign DNA
into an Organism
Cells will multiply and produce desired
gene product
pGlO gene expression vector: Green
Fluorescent Protein
Day 2: Upstream Processing:
Cell Growth and Lysis
Activities
• Transformed cells grown
overnight in selected media
are separated from media
by centrifugation
• Media is removed and
packed cells are lysed
• Homogenate is centrifuged
• Supernatant with pGLO
protein is retained for
downstream processing
Outcomes
• Multiplication of cells
• Initial separation of
fluorescent protein from
cell homogenate
Day 3: Downstream Processing:
Purification of Green Fluorescent Protein by
Chromatography
Activities
• Separation of product by
Different types of
Chromatography
–
–
–
–
Size exclusion
Hydrophobic Interaction
Cationic Interaction
Anionic Interaction
Outcomes
• Fractions with green
fluorescent protein will
glow and be selected
for analysis
• Understand concept of
chromatography as
selective interactions of
compounds with matrix
Isolation and Purification of Green
Fluorescent Protein
Transformed
cells
Test
tubes
Fraction
number
#1
#2
#3
Courtesy of Bio-Rad
Separation of Proteins
by Column Chromatograph
 Separation by size
– Size exclusion chromatography
 Separation by hydrophobic characteristics
– Water loving vs water hating environments
 Separation by ionic charge
– Protein has a positive charge & attaches to
negative matrix on column: Cation Exchange
– Protein has a negative charge & attaches to
positive matrix on column: Anion Exchange
Size Exclusion Chromatography
Red molecule =10^6 daltons - Blue molecule = 600 daltons
How Size Exclusion Works
 Molecular size of molecule will separate two
or more molecules
 Large molecules can not go into a bead of a
certain size and flows quickly through a
column
 Small molecules enter into a bead and flows
slowing through a column.
 Size of two different molecules are separated
Hydrophobic Interaction
Chromatography
H+
H+
H+
H+
Hi Salt
High salt:
Low salt
2M
(NH4)2SO4
Wash buffer:
1.3 M
(NH4)2SO4
Elution
buffer:
10 mM
Tris
Ion Exchange Chromatography
• Proteins bind to
opposite charges on
the matrix
• An example of ion
Exchange
• Addition of
increasing
Salt/pH should
release proteins
Cation Exchange vs Anion exchange
 How proteins are attracted to ion exchangers
Day 4: Quality Control
Activities
• Chromatography fractions
prepared for
electrophoresis
• Electrophoresis Box is
assembled with PAGE gel
• Samples of chromatography
fractions are added to PAGE
gel and ran for 30 minutes
• Gels stained and viewed
Outcomes
• Analysis of protein samples
by observation on light box
determines protein purity
• Standard molecular weight
markers indicate size of
protein
• Verification of mol wt by
comparison with standard
proteins and number of
proteins in a single sample
Quality Control
Analysis of Column Fractions
 Isolated fractions using Ion Exchange
Chromatography are then analyzed
 Electrophoresis by SDS PAGE of fractions
collected
Day 5. Discovery Research and
FDA approval process
Questions
• Discovery Science in Drug
development
• Good Manufacturing
Practices and the FDA
• Initial Biomanufacturing
Process Development
• Scale up to full production
of biologic /clinical trials
Answers
• Visit local biomanufacturing
plants
• Discuss importance of
documentation in the
workplace ie. SOP and Batch
Records
• Invite industry
representatives to speak to
the regulated workplace
Bio-Rad Partnership
Northeast Biomanufacturing Center and Collaborative
(NBC2) is partnered with Bio-Rad:

Provides teachers with engaging hands-on
biomanufacturing education that is easily
accessible

Introduction to:
– metrology,
– production (upstream and downstream processing)
– quality control biochemistry and clinical trials.
These modules can be brought directly into the classroom
Virtual Chromatography
http://www.Atelearning.com/BioChrom
HTTP
Useful Information for
Biomanufacturing
 http://www.Biomanufacturing.org
 http://www.Biomanonline.org
 http://www.Bio-link.org
Background Resources:
“Development of Biotechnology Curriculum for the Biomanufacturing
Industry”, Robert McKown, and George L. Coffman, May/June 2002,
Pharmaceutical Engineering pages 1-6.
“ Introduction to Biomanufacturing a Global Biomanufacturing Curriculum”
Northeast Biomanufacturing Center & Collaborative (NBC2) 2011,
publisher :Lulu.com