How to Start a Biotechnology Program

1. How to Start a BiotechnologyProgram Mary Jane Kurtz, Ph.D NBC2 Consultant mjkurtz@biomanufacturing.org

2. 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

3. 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

4. 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

5. 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

6. 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

7. 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

8. 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

9. 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

11. Student Success Stories Students completing courses with certificate or Associate degrees in Biomanufacturing: www.biomanufacturing.org http://www.biomanufacturing.org/

12. 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

13. 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

14. 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.

15. 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

16. Intro to Biotechnology for High School/CC

17. Biotechnology Curriculum • Associate Degree or Certificate Program Resources: Introduction to Biomanufacturing: Global Biomanufacturing Lab manual NBC2, atwww.biomanufacturing.org Introduction to Biomanufacturing: Textbook, NBC2, at www.biomanufacturing.org Introduction to Biotechnology Lab Manual ATE central: Linnea Fletcher, et alISBN BITC1311006

18. Techniques in Biotechnologyfor Community College

19. Case study of Two Different Programs College Degree High School or Certificate 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 • 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

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

21. 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

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

23. $Protein is Cash$ Local Teachers Workshops Biomanonline.org

24. Day 1 : Metrology/Instrumentation Activities Outcomes Introduction of Good Manu-facturing Practices SOP and documentation Confidence in measurements by instru- mentation • Calibration of top balance • Verification of pipet performance • Pipetman Challenge

25. Metrology

26. Day 2 Transformation of Bacteria Activities 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 • 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

27. 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

28. Results of Inserting Foreign DNA into an Organism Cells will multiply and produce desired gene product pGlO gene expression vector: Green Fluorescent Protein

29. Day 2: Upstream Processing: Cell Growth and Lysis Activities Outcomes Multiplication of cells Initial separation of fluorescent protein from cell homogenate • 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

30. Day 3: Downstream Processing: Purification of Green Fluorescent Protein by Chromatography Activities Outcomes Fractions with green fluorescent protein will glow and be selected for analysis Understand concept of chromatography as selective interactions of compounds with matrix • Separation of product by Different types of Chromatography • Size exclusion • Hydrophobic Interaction • Cationic Interaction • Anionic Interaction

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

32. 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

33. Size Exclusion Chromatography Red molecule =10^6 daltons - Blue molecule = 600 daltons

34. 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

35. 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

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

37. Cation Exchange vs Anion exchange • How proteins are attracted to ion exchangers

38. Day 4: Quality Control Activities 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 • 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

39. Quality ControlAnalysis of Column Fractions • Isolated fractions using Ion Exchange Chromatography are then analyzed • Electrophoresis by SDS PAGE of fractions collected

41. Day 5. Discovery Research and FDA approval process Questions 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 • Discovery Science in Drug development • Good Manufacturing Practices and the FDA • Initial Biomanufacturing Process Development • Scale up to full production of biologic /clinical trials

42. 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

43. Virtual Chromatography http://www.Atelearning.com/BioChrom http

44. 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