How to Start a Biotechnology Program

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How to Start a BiotechnologyProgram

• Mary Jane Kurtz, Ph.D NBC2 Consultant
• mjkurtz_at_biomanufacturing.org

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Why Biotechnology?A number of good reasons

• 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

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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

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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

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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

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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

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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

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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

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Ten Technician Jobs Anchor Ten Biomanufacturing
Departments

• Facilities/Metrology
• Validation
• Environmental Health and Safety (EHS)
• QA
• Upstream Processing
• Downstream Processing
• QC Microbiology
• QC Biochemistry
• Process Development

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RESEARCH DEVELOPMENT(pre-clinical) Discovery Research OPERATIONS Process development, Manufacturing Production QUALITY Quality Control Assurance CLINICAL RESEARCH Clinical Research Regulatory Affairs
Discovery Research Senior Scientist Scientist III,II,I Research Associate Process Development Director Supervisor Process Development Technician Quality Control (QC) Clinical Research Clinical Research Manager
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 degrees 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
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Student Success Stories

• Students completing courses with certificate or
• Associate degrees in Biomanufacturing
• www.biomanufacturing.org
• http//www.biomanufacturing.org/

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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

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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
  BiotechnologyTextbook and Laboratory Reference
• Lisa Seidman et al, Prentice Hall, ISBN
  013-795535-9

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Align Curriculum withProgram 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.

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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
Assistant Professor
Staff Scientist
Independent Investigator
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Intro to Biotechnology for High School/CC
1 Introduction to Biotechnology Use of bacteria, enzymes and fermentation by yeast
2 Metrology Validation of Pipets and Calibration SOP s Documentation of Testing
3 Basic Microbiology Unit 1 Basic Aseptic Technique and Testing of Bioload in Biomanufacturing Areas Gowning up
4 Basic Microbiology Identification of Bacterial Types Unit 2 Identify of Microbe Morphology and Electrophoresis
5 Transformation of Cells With DNA PGlO transformation of Bacteria
6 Spectrophotometry Protein determination of samples By Bradford and UV analysis
7 Genetic Transformation of Bacteria E. coli transformation with pGLO plasmid
8 The Rain ForestHow do we find new pharmaceuticals ? Growth of pGlo transformed bacteria and isolation of the p-GLO product
9 Proteins and their purification Isolation of casein from milk Using solubility and filtration techniques
10 Good Manufacturing Practices Documentation GMP activity with popcorn And use of Documentation
11 Chromatographic separation and how it works Use of ion exchange and hydrophobic columns in Isolating GFP
12 QC Sampling of Product to Determine its Purity Polyacrylamide Gel Electrophoresis
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Biotechnology Curriculum

• 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

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Techniques in Biotechnologyfor Community College
Week Lecture and Topic Laboratory
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 by Ion Exchange Purification of LDH using a IEX column
5 Identification of Proteins Concentration and Electrophoretic Techniques
6 High Pressure Chromatography Quality control
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Case study of Two Different Programs

• College Degree

• High School or Certificate

• 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

• Entry level math is pre-algebra 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

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High School Curriculum
A Workshop to Instruct Teachers and Students
in Biomanufacturing and the Bioeconomy
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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

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• Metrology
• Transformation of Bacteria
• Upstream Processing
• Downstream Processing
• Quantitative Analysis
• Discovery Research

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Protein is Cash Local Teachers Workshops
Biomanonline.org
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Day 1 Metrology/Instrumentation

• Activities

• Outcomes

• Calibration of top balance
• Verification of
• pipet performance
• Pipetman Challenge

• Introduction of Good Manu-facturing Practices
• SOP and documentation
• Confidence in measurements by instru-
• mentation

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Metrology
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Day 2 Transformation of Bacteria

• Activities

• Outcomes

• 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

• 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

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Day 2 E.coli Transformation with pGLO plasmid
Ampicillin resistance gene (Ampr) and target
gene on bacterial plasmid
Only E. coli containing plasmid survive on
Ampicillin plates
Cell division
Transformation mixture is plated on to agar plate
containing Ampicillin
Bacterial clones
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Results of Inserting Foreign DNA into an Organism

• 
  

• Cells will multiply and produce desired gene
  product
• pGlO gene expression vector Green Fluorescent
  Protein

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Day 2 Upstream Processing Cell
Growth and Lysis

• Activities

• Outcomes

• 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

• Multiplication of cells
• Initial separation of
• fluorescent protein from
• cell homogenate

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Day 3 Downstream Processing Purification of
Green Fluorescent Protein by Chromatography

• Activities

• Outcomes

• Separation of product by Different types of
  Chromatography
• Size exclusion
• Hydrophobic Interaction
• Cationic Interaction
• Anionic Interaction

• Fractions with green fluorescent protein will
  glow and be selected
• for analysis
• Understand concept of chromatography as selective
  interactions of compounds with matrix

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Isolation and Purification of Green Fluorescent
Protein
Transformed cells
Test tubes
1 2 3
Fraction number
Courtesy of Bio-Rad
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Separation of Proteinsby 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

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Size Exclusion Chromatography
Red molecule 106 daltons -
Blue molecule 600 daltons
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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

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Hydrophobic Interaction Chromatography

• Low salt

High salt 2 M (NH4)2SO4
H
H
H
H
Wash buffer 1.3 M (NH4)2SO4
Hi Salt
Elution buffer 10 mM Tris
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Ion Exchange Chromatography

• Proteins bind to opposite charges on the matrix
• An example of ion Exchange
• Addition of increasing
• Salt/pH should release proteins

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Cation Exchange vs Anion exchange

• How proteins are attracted to ion exchangers

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Day 4 Quality Control

• Activities

• Outcomes

• 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

• 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

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Quality ControlAnalysis of Column Fractions

• Isolated fractions using Ion Exchange
  Chromatography are then analyzed
• Electrophoresis by SDS PAGE of fractions
  collected

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Day 5. Discovery Research and FDA approval
process

• Questions

• Answers

• Discovery Science in Drug development
• Good Manufacturing Practices and the FDA
• Initial Biomanufacturing
• Process Development
• Scale up to full production
• of biologic /clinical trials

• 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

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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

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http

• Virtual Chromatography
• http//www.Atelearning.com/BioChrom

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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