Northern New England Hub at New Hampshire Community Technical College

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Northern New England Hub at New Hampshire
Community Technical College
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N. New England Learning Community (NHCTC members)

• Sonia Wallman, Director of Biotechnology
• Deb Audino, Biomanufacturing Faculty
• Robert OBrien, Biomanufacturing Facility Manager
• Tim Dubuque, NBC2 Webmaster/Multimedia Guru
• David Miller, IST Faculty

• Kari Britt, Biomanufacturing Intern
• Hope Townes, Biomanufacturing Intern
• Laura Tyson, Biomanufacturing Intern

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N. New England Learning Community

• Lonza, Portsmouth, NH
• Glycofi, Lebanon, NH
• Stryker, Lebanon, NH
• Bentley Pharmaceutical, Exeter, NH
• Bio-Concept, Derry, NH
• Wyeth, Andover, MA
• Biotecture Design, Meredith, NH
• PFI/Parsons Engineering
• Pall Euroflow, Portsmouth, NH
• Bioprocessing, Scarborough, ME
• Wunderlick-Malec, Portsmouth
• TolerRX, Inc., Cambridge, MA
• NH Biotech Council
• Rockingham Economic Development Council

• Ann Gnagey, Biotech Coordinator, Vermont
  Technical College
• P.T. Vasudevan, Chemical Engineering, University
  of NH
• Carolyn Kelley, Biotech Coordinator, Seacoast
  School of Technology
• Ken Franson, Kingswood Regional HS, Wolfeboro, NH
• Rich Parent, Biotech Coordinator, Milford High
  School, Milford, NH

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N. New England Learning Community Job Foci

• Metrology coordinated by Robert OBrien, NHCTC
• Maintains, tests, troubleshoots and repairs a
  variety of circuits, components, analytical
  equipment and instrumentation. Calibrates
  instrumentation and performs validation studies.
  Specifies and requests purchase of components.
  Analyzes results and may develop test
  specifications and electrical schematics.
  Performs continuous monitoring of equipment
  status, condition and location. Prepares required
  documentation for the recording and notification
  of events and changes related to equipment such
  as calibration certificates, deviations, out of
  tolerances and installation reports. Knowledge
  and understanding of measurement parameters and
  experience working in a GMP environment is
  preferred.
• Process Development coordinated by Deb Audino,
  NHCTC
• Responsible for evaluating and scaling-up
  manufacturing processes in order to improve
  product yield and reduce overall costs of
  production. Executes small-to-medium scale
  production work, which may involve cell culture,
  fermentation, purification, chromatography, and
  developing QC protocols. Additionally, assists
  with maintenance of production equipment. May
  research and implement new methods and
  technologies to enhance operations and may assist
  in validation of production processes.
• Facilities coordinated by Sonia Wallman, NHCTC
• Performs monitoring, repair, and preventive
  maintenance activities on critical systems and
  facility equipment. Troubleshoots, installs, and
  modernizes new and existing systems, including
  refrigeration equipment, water systems, HVAC
  systems, and electrical systems. Documents
  repairs, adjustments, and replacement of
  equipment and/or components per GMP standards.
  May also provide input and corrections to SOPs
  and assist engineering in the evaluation of new
  equipment or technology. Knowledge of major
  trades, such as carpentry, electrical, plumbing,
  and HVAC refrigeration, including the ability to
  interpret blueprints, technical manuals, and
  specifications, is required.

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Calibration Metrology

• System approach
• Monitor Verify Validate
• Periodic maintenance
• Periodic calibration
• SOP
• Documentation
• Traceability
• The Real Problem
• Wear
• Misuse
• Hands on component
• Calibration of pH meter
• Calibration of the balance scale
• Calibration of pipette
• Calibration Air sampler

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Calibration Metrology

• Features of a calibration management system CMS
• Identifies instruments that can be calibrated
• Determines calibration requirements for
  instruments
• Requirements for a CMS
• Instrument requirements unique identification
• A recorded history and current calibration status
• Use appropriate for the function of the
  instrument
• Calibration standards and test equipment
• Standards Organizations
• ISO - International Origination for
  Standardization
• ANSI - American National Standards Institute
• UPC - United States Pharmacopeia

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Calibration Metrology

• Training
• Ensure that all personnel involved with
  calibration are properly trained
• Training records must be maintained
• Change Control
• Updates and refinements of the calibration system
  must be done in a methodical and documented
  manner
• Electronic Records
• Management of electronic records for paperless
  calibration operations
• Auditing
• Periodic audits assure that the calibration plan
  is being followed

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Calibration Metrology

• CALIBRATION PHYSICS AND INSTRUMENTS
• Aseptic
• Volume
• Temperature
• Electrical
• Measurement
• If a parameter is not measured, it cannot be
  controlled
• Statistics
• Method for handling data and drawing conclusions
• PM
• Ensure correct results are capable of being
  obtained
• Aseptic Practices
• Used while performing calibration to avoid
  inadvertently cross-contaminating

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Calibration Metrology

• Equipment
• Autoclave
• pH Meter
• Pipette
• Temperature
• IR Thermometer
• Transducers/ RTD
• Thermocouples
• Freezers, Refrigerators
• Hot plate
• Incubator
• Pressure Calibration Meter
• Biological Safety Cabinet
• Microscope
• Humidity
• Tachometer
• Centrifuge
• Digital Multimeter

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Calibration and Monitoring
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Calibration Metrology

• Lecture and Hands on Lab components presented at
  Bio-man July 2006 129 Slide power point
• 20 Registered Participants
• Participants calibrated a balance utilizing the
  calibration SOP
• Validation / Monitoring of the autoclave
• Participants performed the biological indicator
  SOP
• The students were able to observe a previously
  biological indicator that had been incubated the
  required time and temperature
• Equipment purchased on a DOL grant was
  incorporated into the development of the
  Calibration Metrology for the NSF grant

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Calibration Metrology

• Future Areas of development
• Additional SOPs for PM need to be completed prior
  to calibration SOPS can be developed for
  equipment
• The need for additional equipment purchases will
  need to be evaluated
• One area utilized this year was behavioral Video
  that was very beneficial for an participant to
  review video of a process being performed
• This behavioral video aspect is being looked at
  to see if it can be incorporated into calibration
  metrology
• Possibility of development of an data base in
  Microsoft Access

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Process DevelopmentOverview
Independent Student Projects Designed to
Create Curriculum for Process Development
Modules UPSTREAM Modules Comparison of Cell
Types Comparison of Medias Comparison of Culture
Vessels DOWNSTREAM Modules Increase the
purity of proteins Design purification
schemes Scale up purification
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Process DevelopmentStudent Externs-Past
Independent Projects
Purification of tPA Improve the purification of
tPA by adding an affinity chromatography step
Yield Function Ease of
purification
Kari Britt
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Process DevelopmentStudent Externs-Current
Independent Projects

• Compiling the Pichia pastoris HSA Core Production
  System

This process yields an ample supply of HSA which
can be used in a process development focusing on
scaling up purification to a 1L column
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Process DevelopmentStudent Externs-Current
Independent Projects
Cloning tPA into Pichia Cells Comparison of tPA
production in CHO vs Pichia cells Yield
Function Ease of purification
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Process DevelopmentStudent Externs-Current
Independent Projects

• Make competent cells
• Transform Invitrogen plasmids into competent
  cells
• Isolate plasmids
• Make cDNA library from CHO cells expressing tPA
• Add EcoR1 linker
• Cut plasmids with EcoR1
• Ligate cDNA fragments and plasmids
• Transform plasmids into competent cells
• Screen for correct plasmid using colony
  hybridization
• Sequence positive clones for mutations
• Transform plasmids into Pichia
• Screen for tPA production in Pichia cells

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Process DevelopmentStudent Externs-Current
Independent Projects
Production of Herceptin Antibodies are the
fastest growing type of biopharmaceutical
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Process DevelopmentStudent Externs-Current
Independent Projects

• Learn Mammalian Cell Culture with CHO Cells
  Expressing tPA
• Aseptic Technique
• Counting cells and determining viability with
  Tyrpan Blue Assay
• Scaling up the culture
• Grow the Hybridoma Cells Expressing Herceptin
• Purify Herceptin using a Protein A Affinity
  Column

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Process DevelopmentStudent Externs-Future
Independent Projects
Scale Up the Purification of HSA
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Process DevelopmentStudent Externs-Future
Independent Projects

• Evaluate Wave Disposable Bioreactor with
• Spinner Flasks and Bioreactors

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Rationale for the NH Pilot Plant

• To support a case study on the design,
  construction, and validation of a
  biomanufacturing facility to be used in the NBC2
  global Biomanufacturing course
• To provide students with visual access to a
  biopharmaceutical manufacturing facility
• To give some graduates of NHCTCs biotechnology
  program actual experience in biopharmaceutical
  manufacturing
• To provide some experienced individuals for
  biomanufacturing companies
• To support the development of biopharmaceuticals
  for eventual commercial manufacture

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Rationale for the NH Pilot Plant

• Rationale comes directly from NHCTCs involvement
  with biomanufacturing education and training and
  our desire to expand our Virtual WorkplaceTM
  concept to include familiarity with the
  operations of a cGMP Pilot Plant.
• Design a cGMP Pilot Plant that uses easily
  validated, completely disposable equipment and
  supplies for upstream and downstream processing
  for ease of cGMP compliance.
• Document design and construction of the cGMP
  Pilot Plant to be used in the NBC2 global
  curriculum addressing a general understanding of
  the construction and operation of a cGMP
  biopharmaceutical manufacturing facility.

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NH Pilot Plant Funding

• Work with county and federal agencies to fund the
  construction of the Pilot Plant, a process
  development lab and offices for incubatees
• Work with John Rafuse, AIA and MEP person to
  design the Pilot Plant
• Work with NHCTC-Stratham/Pease architects in the
  construction of the Pilot Plant
• Work with industry partners to equip a fully
  disposable Pilot Plant
• Pall Filtron will provide TFF equipment
• Fisher Scientific will provide some disposable
  supplies and equipment for the Pilot Plant and
  equipment for QC and process development
• Nalge Nunc may provide a variety of disposable
  supplies
• Looking for other industry partners to provide
  downstream processing disposable equipment and
  supplies

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Pilot Plant Flow Diagram
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Pilot Plant Walls and Doors Plan
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Pilot Plant View from Outside
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Pilot Plant Interior Shot
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NHCTC as the Center for NBC2 Activities

• NHCTC is the site for
• The annual NBC2 NVC meeting in March
• The annual 5 day BIOMAN conference in July or
  August
• Northeast Region of Bio-Link
• Value-added NBC2 activities stemming from NHCTC
  Department of Labor grants, one to build capacity
  for education and training at NHCTC and another
  to develop the infrastructure for biotechnology
  and biomanufacturing education and training in
  New Hampshire

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The NH BET Facility

• A 5,000 square foot facility for biotechnology
  education and training with a wet lab for
  Discovery Research, a Biomanufacturing Suite and
  a Core Facility for shared equipment
• Constructed to resemble a vertically integrated
  biotechnology company with the tools, processes
  and regulatory structure for Discovery Research,
  Process Development and Biomanufacturing.
• Providing education and training in Industrial
  Biotechnology for all levels recent high school
  graduates, UNH students, career changers,
  displaced workers, women returning to work,
  incumbent workers, faculty and teachers

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Biomanufacturing Equipment

• Multiscan ELISA Plate Reader
• Drummond Automatic Pipettors (4)
• Ohaus Balances (4)
• Conductivity Meter
• Orion pH and Conductivity Meters (4)
• Applikon 3 liter process controlled
• Bioreactors (4)
• New Brunswick Bioflo 3000 process
• controlled bioreactor
• Bio-Rad Liquid Chromatography Systems (4)
• Millipore 2L chromatography columns
• Nova 300B analyte analyzer
• MET One laser particle counter
• Millipore air sampler
• Bio Tek Microplate Reader
• Sanyo Dual CO2 Incubator
• Sanyo Top Loading Autoclave
• UVP Digital Photodoc System
• Invitrogen Mini-Cell SDS-PAGE Gel Boxes (4

• Bio-Rad HPLC System
• Forma biological safety cabinet
• Hot Plate/Magnetic Stirrers
• Microfuges
• Rainin Pipetman
• Household (non-defrosting) refrigerator
• Freezers (-20 and -86 degrees C)
• Cellstar dry shaking incubator
• Market Forge autoclave
• Millipore Milli Q water system
• Bellco spinner bottles
• Bellco magnetic stirrers
• Olympus BX40 uv-visible video microscope
• Olympus CK2 inverted microscope
• Leica Microscopes (4)
• Dell laptops with remote observation software for
  Applikon bioreactors, Bio-Rad LC system, Biolog
  system and Microplate Reader (4)
• Applied Biosystems GeneAmp PCR system
• Sigma refrigerated centrifuge
• Sanyo incubator

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Synergistic Activities

• Member of the Advisory Group of the Biotechnology
  Labor Market Initiative, a DOL funded project
  (identify the industry, the jobs and salaries,
  the sources of education, career ladders/latices,
  etc. and build website to communicate this
  information)
• Formation of the Northeast Biomanufacturing
  Institute offering short courses for incumbent
  workers (part of DOL grant that created the
  Center of Expertise in Biomanufacturing, part of
  the National Center for the Biotechnology
  Workforce-www.biotechworkforce.org)
• Northeast Region Director of Bio-Link (the
  Bio-Link webmaster resides here)
• Work with AACC and FAS, Ireland re
  biomanufacturing education and training,
  workforce, and apprenticeships

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Synergistic Activities, continued

• Development of the Registered Apprenticeship in
  Biomanufacturing (part of DOL grant that created
  the Center of Expertise in Biomanufacturing)
  NBC2 involved in Regionalization of the
  Biomanufacturing Apprenticeship program-we are
  looking at the Biomanufacturing Apprenticeship as
  a long-term internship for students (this was not
  listed as a deliverable because we thought it was
  an impossible task).
• NHCTC Biotechnology Strategic Enrollment Program,
  John Grady, Consultant (part of DOL Grant that
  created Center of Expertise in Biomanufacturing)
  Targets Apprentices, NHCTC System CCs, Industry,
  Four Year Colleges and Universities

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Student Profile 1994-2006

• Total number of graduates323
• Total number of graduates at work268
• Total number of graduates at biomanufacturing
  companies182
• Companies employing biomanufacturing graduates
  Lonza-NH, Wyeth-MA, Superior Controls-NH,
  Amgen-RI Amgen-CA, Genzyme-Alston-MA, Marine
  Polymer Technologies-MA, TKT(Shire)-MA,
  Arqule-MA, Bioprocessing-ME, IDEXX-ME,
  Bio-Concept-NH, East Coast Validation-NH,
  Roche-NJ, Formatech-MA, Beckton-Dickinson-MA,
  Igenex-CA, Repligen-MA, Hyaluron-MA, Alerchek-NH,
  GlycoFi (Merck)-NH, Genentech-CA, Valicor-MA,
  Dakota Systems-MA, Millipore-NH, BiogenIdec-MA,
  Genzyme-MA
• Companies currently providing Biomanufacturing
  Apprenticeships Lonza-NH, Bentley
  Pharmaceuticals-NH, NH BET Center-NH