The document provides information about biomanufacturing education and training programs. It discusses the Northeast Biomanufacturing Collaborative and Center's work since 2003 to define the skills, knowledge, and abilities needed by biomanufacturing technicians. It highlights the development of curriculum and instructional materials to educate technicians for careers supporting the bioeconomy. Examples are given of various biomanufacturing processes and the equipment involved in areas like biopharmaceuticals, biofuels, and industrial biotechnology.

1. The Bioeconomy
Bioproducts
Development, Production and Analysis
Education and Training
Skilled Technicians
Needed
Sonia Wallman, PhD
NBC2 and c3bc
Biomanufacturing Consultant
Bio-Link Summer Fellows
Berkeley, CA
June 8, 2016

2. History of Northeast Biomanufacturing
Collaborative and Center
Since 2003, working with biomanufacturing employers to define
the skills, knowledge and abilities (SKAs) needed by technicians to
develop, produce and analyze bioproducts. Starting with
biopharmaceutical production and analysis in 2003, 2007 and
2013; with biofuels in 2013 and finishing up with industrial
biotechnology in 2015 (and 2016), we will synthesized these inputs
from local (and global) employers into a single set of SKAs to
educate and train technicians to support the development and
growth of the bioeconomy. The synthesis of these inputs on what
biotechnicians need to know and do is driving the production of
hands-on and online curriculum and instructional materials,
including laboratory manuals, textbooks and virtual industrial
modules to facilitate deep understanding of what is required by
technicians to biomanufacture bioproducts, creating an
awareness of the many career pathways in biomanufacturing.

3. Biofuels Workforce Summit
May 23, 2013 in Honolulu, HI

4. Bioproducts Upstream Processes
BIOPHARMACEUTICALS,
INDUSTRIAL ENZYMES, GREEN
CHEMICALS, BIOFUELS
• DEFROST CRYOVIAL OF CELLS
FROM WORKING CELL BANK
• INOCULATE MEDIA WITH
CRYOVIAL OF CELLS
• SCALE-UP
• HARVEST
• CENTRIFUGATION
• FILTER MEDIA (DEPTH
FILTRATION AND STERILE
FILTRATION)
• HOLD
BIODIESEL
FEEDSTOCKS: WASTE AND AG
• COLLECT OIL PRODUCTS SUCH
AS WASTE FRY OIL AND PLANT
OILS
• TRANSPORT
• STORE
• HEAT
• FILTER
• HOLD

5. Bioproducts Downstream Processes
BIOPHARMACEUTICALS,
INDUSTRIAL ENZYMES, GREEN
CHEMICALS, BIOENERGY
• DEPTH FILTRATION
• LIQUID COLUMN
CHROMATOGRAPHY (ION
EXCHANGE, AFFINITY,
HYDROPHOBIC
INTERACTION=HIC, SIZE
EXCLUSION=GEL FILTRATION)
• TANGENTIAL FLOW
FILTRATION
• STERILE FILTRATION
• COLLECT BATCH
• FORMULATE, FILL AND FINISH
(for biopharmaceuticals)
• DISTRIBUTE
BIODIESEL
FEEDSTOCKS: WASTE AND AG
• TRANSESTERIFY OIL WITH
ALKALINE CATALYST AND
METHANOL INTO FAME (FATTY
ACID METHYL ESTERS=BIODIESEL)
• SEPARATE INTO FAME-MeOH AND
GLYCEROL-MeOH LAYERS
• DISTILL MeOH FROM BOTH
LAYERS – RECYCLE MeOH
• WATER WASH FAME LAYER AND
FILTER OR DISTILL AT HIGH
VACUUM
• DISTRIBUTE BIODIESEL
• SELL GLYCEROL CRUDE OR
PURIFIED BY HIGH VACUUM
DISTILLATION

6. Pacific Biodiesel (Big Island Biodiesel)
5M gallon/year Fry Oil to Biodiesel Facility
Collected fry oil into heated
tanks, oil strained, placed into
funnel shaped tanks where
sodium hydroxide cleaves
glycerol from triglyceride and
methanol is used to add a
methyl group to the free fatty
acids to produce FAME or fatty
acid methyl esters or biodiesel.
Glycerol to bottom of funnel
and biodiesel is drawn off and
dewatered. BIB follows this by
distillation, producing crystal
clear biodiesel.

7. Vessels and Pumps
Vessels store/contain liquids, solids, or
gasses involved in bioproducts
production, such as bioreactors, tanks,
drums, cylinders, and bins.
Pumps are used to move liquids through
the system, such as into or out of a vessel.
There are many different types of pumps,
but they all use either rotational or
centrifugal force to induce flow in liquids.
Tank holding Water For Injection (WFI) Peristaltic Pump

8. Compressors, Fans and Motors

9. Piping and Valves
Piping is cylindrical tubes made of
varying lengths and materials such as
316L stainless steel, plastics, and iron
that carry materials (usually in liquid or
gas form) from one location to
another.
Valves are devices that are attached to
piping or equipment to control the
flow of liquids or gases. There are a
wide variety of valve types that can
start/stop or throttle (open or close in
increments) the flow. Valves are vital
to process control. They can be
remotely opened or closed by
computers to control variables such as
flow or level.
Piping and valves are the most
common pieces of equipment found in
a biomanufacturing facility.

10. Piping & Instrumentation Diagrams
P&ID
A drawing or diagram
which shows the piping of
the process flow together
with the installed
equipment and
instrumentation, including
computer process
controllers.

11. Biomanufacturing is Process Controlled
Big Island Biodiesel 5M Gallon/Year

12. Quality Control Laboratory
BIOPHARMACEUTICALS,
INDUSTRIAL ENZYMES, GREEN
CHEMICALS, BIOFUELS
BIODIESEL
TriState Biodiesel in Bridgeport, CT
Built Using Recycled Equipment
Lonza Biopharmaceuticals
in Portsmouth, NH

13. Quality Control Testing
BIOPHARMACEUTICALS,
INDUSTRIAL ENZYMES, GREEN
CHEMICALS, BIOFUELS
BIODIESEL
• Cell Count (growth rate)
• Analyate Analysis (metabolism)
• ELISA (protein ID, amount, and
activity)
• SDS PAGE (protein ID and amount)
• TLC (thin layer chromatography)
• HPLC plus detector (protein, lipid,
carbohydrate IDs and amount of
each)
• Stability Testing
• Genome Sequencing
• PCR Identification

15. Scale-Up Using Aseptic Technique
Shake Flask Inoculation using Biosafety Cabinet Class 100

16. Upstream Processing Count Cells

17. Scale-Up to
Stainless Steel Biopharmaceutical Production
Bioreactors

18. Inoculation of Microalgae Seed Bioreactor

19. Scale-Up of Microalgae to
Open Pond Production Solar Bioreactors

20. Sludge
Continuous
Centrifuge
Media Out
Cells + Media In
Harvest Protein Rich Media
by Continuous Centrifugation

21. Harvest by Batch Centrifugation
(1000 liters of Chlorella vulgaris culture)
Process controlled centrifuge to
separate cells (microalgae) from media
Centrifuge drum and fins for
separation of microalgae from media

22. Cellular paste is
collected below
the blades of
the centrifuge
Microalgae are
harvested; media is
reprocessed into
useable water

24. Depth Filtration to Remove Cells

25. Depth Filtration Housings, Piping, Valves

26. Tangential Flow Filtration (TFF) for
Separation and Concentration
Using TFF with the right cut off filters, the biologic of interest
can be separated from other thing
For instance HSA has a molecular weight of 69KD. To make
sure that the protein of interest is retained, a 10KD cut-off
filter is used.
After ultrafiltration, we can diafilter, adding the phosphate
buffer at pH 7.1 that we will also use to equilibrate our
affinity column to prepare it for affinity chromatography of
HSA.

27. Tangential Flow Filtration
Separates and Concentrates

28. TFF for Biopharmaceutical Protein
Concentration or Buffer Exchange

29. TFF for Microalgae Concentration
Photo taken at the ATP3 Large Scale Algal Cultivation, Harvesting and Downstream Processing
Hands-On Training at AZCATi (Arizona Center for Algal Technology and Innovation) in Mesa, AZ
From Open Pond
To Centrifuge

30. Chromatography for Purification and
Quality Control Chemistry of the Protein,
Lipid or Carbohydrate of Interest
Separation Science Used for Downstream Processes
and for Quality Control Chemistry Monitoring of both
Upstream and Downstream Processes
Stationary Media and Moving Buffer System
Result in Competition and Separation

31. Liquid Column Chromatography

32. Large Scale Chromatography Column
Packed with Beads (Media) for Separation

33. Chromatography System Components
• Chromatography Column
and Media (Beads)
• Peristaltic Pump, Pipes
and Valves
• Mixer for Buffers
• (UV) Detector to identify
the product of interest

34. Liquid Column Chromatography Process
PURGE Air from Column using Equilibration
Buffer
PACK Column with Media (e.g. ion exchange, HIC,
affinity or gel filtration media)
EQUILIBRATE Column with Equilibration Buffer
LOAD Column with Protein of Interest in
Equilibration Buffer
WASH Column with Equilibration Buffer
ELUTE Protein of Interest from Column with
Elution Buffer of high or low Salt or pH
REGENERATE Column or Clean and Store (NaOH)

35. Chromatogram
WASH=Flow Through ELUTE=Eluate

36. Skills, Knowledge and Attributes
for Biomanufacturing Technicians
Specific SKAs for Biomanufacturing Time Spent/Importance of Each
Production and Processing 22
Operation and Control 7.6
Quality Control Analysis 5.6
Computers and Electronics 3.9
Operation Monitoring 3.2
Equipment Selection 2.3
Equipment Maintenance 1.3
Repairing 1.1
Engineering and Technology 0.7
Design 0.7
Systems Analysis 0.6
Technology Design 0.1
Installation 0.1

37. Lab Science, Medical Devices and
Biomanufacturing
Common Core Critical Work Functions
• Maintain a Safe and Productive Work Environment
• Provide Routine Facility Support
• Perform Measurements/Tests/Assays
• Comply with Applicable Regulations and Standards
• Manage and Communicate Information
• Perform Mathematical Manipulations

38. Online Biomanufacturing Resource
www.biomanufacturing.org/
CHAPTER 1 INTRODUCTION
ENGINEERING UNIT OVERVIEW
CHAPTER 2 FACILITIES
CHAPTER 3 METROLOGY
CHAPTER 4 VALIDATION
CHAPTER 5 ENVIRONMENTAL, HEALTH,
AND SAFETY (EHS)
CHAPTER 6 OPERATIONAL EXCELLENCE
QUALITY UNIT OVERVIEW
CHAPTER 7 QUALITY ASSURANCE
CHAPTER 8 MICROBIOLOGICAL CONTROL
CHAPTER 9 QUALITY CONTROL
BIOCHEMISTRY
PRODUCTION UNIT OVERVIEW
CHAPTER 10 UPSTREAM PROCESSING
CHAPTER 11 DOWNSTREAM PROCESSING
CHAPTER 12 PROCESS DEVELOPMENT
APPENDIX A MASTER GLOSSARY
Michael Cicio - Today’s
Biologics: From Bench
Top to Bottle.
Tags: Keynote
Speeches, Bioman 2006

39. Virtual Downstream Processing
Monoclonal antibody proteins (or mAbs) are the
single largest class of recombinant biological drugs
to date and represent about a third of the total
biopharmaceutical market. The recent success of
monoclonal antibodies for a wide range of disease
therapies has led to the development of industrial
production operations that manufacture
pharmaceutical-grade mAbs both efficiently and
safely. The following modules introduce a typical
mAb bioprocessing workflow, detailing the
equipment and processes used in biomanufacturing
within a regulated environment.
http://faculty.mc3.edu/downstreamprocessing/stor
y.html

40. NBC2 Biofuels Curriculum – Textbook

41. NBC2 Biofuels Curriculum – Laboratory Manual
Table of Contents
► LAB 1: INTRODUCTION TO BIOFUELS PRODUCTION ........................ 05
► LAB 2: BIOFUEL ENZYME KIT ............................................................. 25
► LAB 3: USING BACTERIA TO PRODUCE BIOHYDROGEN ................... 39
► LAB 4: ESTABLISHING AND SCALING UP MICROALGAE CULTURE
FOR BIOFUELS PRODUCTION SOP ........................................ 55
► LAB 5: CONVERSION OF MICROALGAE LIPIDS TO FATTY ACID
METHYL ESTERS (FAME) SOP.................................................. 59
► LAB 6: MICROALGAE LIPID EXTRACTION AND SEPARATION BY
THIN LAYER CHROMATOGRAPHY SOP.................................... 63
► LAB 7: USING ALGAE TO PRODUCE BIODIESEL................................ 69
► LAB 8: EXTRACTION OF ALGAL OIL .................................................. 83
► LAB 9: PRODUCTION OF BIODIESEL BY THE FOLCH METHOD ......... 89
► LAB 10: BIODIESEL ANALYSIS USING HPLC ........................................ 103
Biofuels Productio
n
and Anal ysis
Laboratory Manual
Primary Author: Elmar Schmid, Ph.D.
Associate Professor, Biotechnology Department
Miracosta College, Oceanside, California
2nd
Editio
n
Apr il 2014
Part One

42. Career Pathway Data Collection:
Job Titles of Some Program Graduates
• Environmental Monitoring Technician
• Laboratory Automation Scientist
• Laboratory Technician
• Manufacturing Associate I
• Manufacturing Tech I
• Manufacturing Tech 2
• Process Improvement Leader
• Quality Assurance Associate
• Quality Analyst
• Quality Control Analyst
• Quality Operations Product Release Coordinator l
• Research Associate
• Research Scientist
• Research Technician
• Senior Laboratory Technician
• Senior Scientist
• Technical Writer
• Technical Writer/Scientist III