Biomanufacture of Cell and Gene Therapies

Cell-based and gene therapy products are growing and becoming a major component of the biotechnology industry. A recent initiative funded by NIIMBL brought together a collaborative of six partner institutions and their regional CGT industry partners. The goals of the WEBET project, or Workforce Expansion in Biomanufacturing Emerging Technologies, are to research the workforce needs of the sector,  identify the technical skills and knowledge needed to develop, manufacture, and test these advanced therapies and finally to create hands-on curriculum that will produce well trained graduates ready to join this important biotechnology sector.

1. REPORTS

1. FACULTY PROFESSIONAL DEVELOPMENT
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3. On December 3, 2021, and January 24, 2022, InnovATEBio hosted WEBET Train-the-Trainer Webinars- view presentations here:                                                                  

CELL AND GENE THERAPY TRAIN THE TRAINER WEBINAR 1  (Recorded December 3, 2021)  Presentations include: CGT labor market research outcomes, technical skill standards for CGT technicians, course requirements, and newly developed lab-based curriculum on AAV production in HEK 293 cells.                                                             

CELL AND GENE THERAPY TRAIN THE TRAINER WEBINAR 2 ( Recorded January 24, 2022)   Presentations include: Skill Standards and Curriculum for Training Cell and Gene Therapy Technicians, Regulatory Considerations for CGT, CGT Analytics, and newly developed lab-based curriculum on AAV production analytics and AAV transduction evalutation by flow cytometry, immunofluorescence and western blot.

• Presenters and affiliations: 
• Dr. Maggie Bryans Montgomery County Community College
• Professor Jim Dekloe Solano Community College
• Russ Read Forsyth Technical Community College
• Drs. Barbara Juncosa and Dominique Ingato MiraCosta College
• Professor Isso Bayala Quincy College

CELL AND GENE THERAPY CURRICULUM

Small Scale AAV Production in HEK293 cells

1. Upstream Processing 
2. AAV-GFP Production in HEK 293F Cells - Upstream Processing Flow Chart                                        
3. SOP: Resuscitation and Culture of Viral Production Cells 2.0 - HEK293F
4. SOP: Cryopreservation of Suspension Viral Production Cells 2.0 - HEK293F

5. SOP: Isolation of pAAV Production Plasmids Using ZymoPURE II Plasmid Maxiprep Kit

6. SOP: Transfection and Harvest of Viral Production Cells (VPC) 2.0-HEK293F for rAAV-CMV-GFP Viral Particle Production 

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8. Downstream Processing

9. SOP: Purification of AAV GFP Viral Particles by Affinity Chromatography

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• Quality Control 
• SOP: Analysis of rAAV Purity by SDS PAGE and Silver Stain                                                                  SOP: Transduction of CHO K1 Cells with Chromatography Purified rAAV GFP Viral Particles              SOP: Viral Gene Delivery and Evaluation Using Flow Cytometry
• SOP: Transduction of CHO K1 Cells with Commercial Viral Vector AAV/DJ-CMV-GFP
• SOP: AAV2 GFP Transduction of CHO K1 Cells: Verification of Transgene Expression by Western Blot 
• SOP: Quantitation of Viral Genome Titer of Affinity Chromatography Purified Adenovirus Associated Viral Particles by qPCR
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• Aseptic Processing
• SOP: Aseptic Processing: Media Transfer Using Tube Welding
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Equipment Required for Training Cell and Gene Therapy Technicians

AAV Vector Manufacturing: Challenges and Opportunities

The Alliance for Regenerative Medicine (ARM) and the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) released Project A-Gene and Project A-Cell. These were multistakeholder collaborations to incorporate Quality by Design (QbD) principles into a manufacturing case study of a viral vector commonly used in gene therapies and a cell-based therapy. View or download a pdf of the case studies below.

1. Project A-Gene
2. Project A-Cell