Production Assistance for Cellular Therapies ‐ PACT
Accelerating Your Cell Therapy
Tuesday March 14, 201712:00 12:00 –– 1:15 PM ET1:15 PM ET
NHLBI – PACT history, scope and application criteria
PACT facilities ‐ Product manufacturing capabilities• David McKenna - University of Minnesota, Molecular and Cellular
Therapeutics• Aisha Khan - Interdisciplinary Stem Cell Institute Cellular
Today’s Web SeminarToday’s Web Seminar
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• Aisha Khan Interdisciplinary Stem Cell Institute CellularManufacturing Program, University of Miami
• Joseph Gold - Center for Biomedicine and Genetics, City of Hope• Linda Kelley - Moffitt Cancer Center• Adrian Gee - Center for Cell and Gene Therapy, Baylor College of
Medicine
Emmes ‐ Application Process
PACT PACT ‐‐ OverviewOverview
PACT Goal – advance cellular therapeuticsfrom the laboratory bench into clinicaltrialsEd ti l P Educational Programs
Resource Center
PACT Services
PACT Website
https://www.pactgroup.net
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PACT Program Life CyclePACT Program Life Cycle
Began September 2003• Cell product manufacturing for clinical trials• 3 cell processing facilities
Renewed January 15, 2010
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• Scope expanded to include translational services• 5 cell processing facilities
Renewed in July 2016• Revised MissionProvide assistance for cellular therapy translational research and the manufacture of cellular therapy products
PACT Organizational StructurePACT Organizational Structure
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PACTPACT‐‐ Requests for Services Requests for Services
PACT will review requests for translational services inthe context of what is required in a product development plan and with an end goal of an eventual IND submission.
PACT Cell Processing Facilities provide the requestedproduct development services and cellmanufacturing for approved projects.
PACT does not provide direct funding to investigators.
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PACTPACT
Cell products that aid in the repair and regeneration ofdamaged and diseased tissues, organs, and biologic systems
Preclinical studies including basic and translational work
Beginning after preclinical therapeutic proof‐of‐concept studies though to validation studies immediately prior toPhase I clinical trial
Products and Services of programmatic interest to NHLBIor with support from other NIH Institutes
Proposals possessing procedural advancements tofurther foster and standardize cell therapies
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Cardiac repair and disease
Lung repair and disease
Hematologic repair and disease (non‐malignant)
H t i ti ll t l t ti i l di ft
NHLBI Scientific InterestsNHLBI Scientific Interests
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Hematopoietic cell transplantation; including graft manipulation and complications (GVHD; graft failure,infections)
Red blood cell, platelet, or white blood cell productsfor transfusion
For more information on PACT Application: Scope Criteria visit www.pactgroup.net
Other NIH Institutes and Federal partners have theability to support investigators through the PACT program
Determined on a case‐by‐case basis
Projects outside of NHLBI InterestProjects outside of NHLBI Interest
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Determined on a case by case basis
Investigators with projects that are outside of NHLBI scientific interests are strongly encouraged to discuss the possible use of PACT with their NIH or Federal Agency program officer and with the PACT NHLBI Program Director prior to starting an application
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ApplicationsApplications
Products and Services
• Supporting activities that will bring proof ofconcept discoveries into the translational development stage.
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p g• Scale‐up
• Optimizing manufacturing processes
• Pre‐clinical process development
• SOP development
• CMC section development
• Cell product manufacturing for relevant animal model studies and other IND‐enabling studies.
Also see https://www.pactgroup.net/content/pact‐services
NHLBI Contact InformationNHLBI Contact Information
Lisbeth Welniak or Traci Heath MondoroPhone: 301‐435‐0065Email: [email protected]
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@ g
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Molecular & Cellular Therapeutics
David H. McKenna, M.D.PACT WebinarMarch 14, 2017
Molecular & Cellular Therapeutics
www.mct.umn.edu
• Class 10,000 clean suites
• Single pass air, isolated differential air pressure rooms, negatively pressured to adjoining rooms (can be adjusted)
• Rooms terminal HEPA‐filtered air tested at1 000 10 000
MCT ‐ Production Space
www.mct.umn.edu
1,000‐10,000 particles/ft3, 20‐70 airchanges per hour
• Production rooms are constructed with epoxy liquid flooring coved base, reinforced resinous wall surfacing, and epoxy painted veneer plaster ceilings
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Recent & Current CT ProjectsProduct Type Procedure Technology
Allo NK (PB) Short-term cx/long-term cx
IM selection, IL-2/IL-15 activation, expansion
Allo Treg (UCB) Long-term cx IM selection, 3/28 or K562 (KT64.86) expansion
Allo MSC (BM) Long-term cx MNC (density gradient), cell factories
Allo UCB expansion Long-term cx IM selection, expansion
Allo UCB homing Incubation Short-term C3a incubation
Auto marrow MNC MNC (automated) MNC (density gradient)
Dendritic cell vaccine [glioblastoma l if (GBM)]
Mid-term cx IM selection, flasksmultiforme (GBM)]
Dendritic cell/tumor cell hybrid vaccine (multiple myeloma)
Mid-term cx PEG-driven fusion
GBM vaccine (lysate) Long-term cx, nebulization
MCB expansion
Allo iTreg (PB) Long-term cx IM selection, KT64.86 expansion
iPS cell banking platform Long-term cx Sendai virus
Virus-specific T cells (CMV, EBV, Adeno, etc.)
Cytokine-capture technology
IM selection
Right ventricular outflow tract (decellularized tissue tube)
Long-term cx Fibrin-based matrix, tube mold
MethodsPB‐Derived NK Cells
anti‐CD3/19
www.mct.umn.edu
Overnight:• XVIVO-15• IL-2• AB serumLot release testing
•CD3-/CD56+•CD3+•Viability•Endotoxin•Gram stain
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MethodsUCB‐Derived Treg Cells
anti‐CD25
www.mct.umn.edu
Day 0:1 x 106 cells/mL in:X-VIVO 15 10% AB serumL-glutaminen-acetylcysteineanti-CD3/CD28 beads (UPenn)
Day 3:IL-2 at 300 units/mL
Day 5-14:0.5 x 106 cells/mL
Day 15-18(+/-1):1 x 106 cells/mL
•Bead removal•Lot release testing
MSC Manufacturing
www.mct.umn.edu
• Trilineage assays• KGF assay
Manufacturing of Induced Pluripotent stem cells (iPSCs)
www.mct.umn.edu
James Dutton, PhD, Jakub Tolar, MD
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Tissue Engineering
• Decellularized tissuetube from humanneonatal fibroblasts forpediatric rightventricular outflow
Tubular Tissue Engineered Valve
www.mct.umn.edu
ventricular outflowreconstruction (RVOT).
• Cells are entrapped onfibrin gel tube
Physiological compliance
High burst strength
Completely biological
Allograft (acelluar)
Syedain et al 2014, Tissue Engr AMeier et al 2014, J Cardiovas TranSyedain et al 2013, Ann Biomed EngrSyedain et al 2010, Biomaterials
Final product is a decellularized off-the-shelftissue-engineered valve
R Tranquillo, Z Syedain
Collaborators/PIs:Bruce Blazar Jeff MillerClaudio Brunstein Zeeshan SyedainSarah Cooley Jakub TolarJames Dutton Bob TranquilloMichael Matthay John Wagner
Thank you!
CT Lab:Diane Kadidlo
Kristen ReynaMichelle Lucio
www.mct.umn.edu
Darin SumstadLisa VanOrsowNancy BostromSheryl AdamsNancy ColeyCindy StanawayLien LeMolly GroweAnh DoStacy LinnAlemayu Emiru
Michelle LucioJulie LaTour
MCT QA:Fran RabeAlison JakubekMaria Opitz& Materials Management/Administrative & Support Staff
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Interdisciplinary Stem Cell InstituteUniversity of Miami
Cell Processing Facility (CPF)
03/14/2017
Aisha Khan, M.Sc, M.B.A
Executive Director of Laboratory Operations
Interdisciplinary Stem Cell Institute
The Miller School of Medicine, University of Miami
Interdisciplinary Stem Cell Institute
• Serves as a manufacturing resource for the various clinical programs
• Facilitates translational research
• Provides regulatory and scale-up expertise to bridge the gap between basic and clinical research applications
• Provides services throughout the IND application
Cell Processing FacilityIntroductionIntroduction
development process, from the proof-of-principle and pre-clinical testing to clinical trial phase.
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Services regional and national research efforts in reparative/regenerative medicine
Equipped to do Phase I, II and III clinical trials
Facility CredentialsFacility Credentials
TRANSLATIONAL CELL
PROCESSING
FDA registered
Serves as a national resource for translational research in the area of cellular therapy
Equipped to manufacture licensed cellular products for new therapies and tissue‐engineered end‐products.
Biomedical Research Building, 9th floor(12,931.28 sq. ft., 85 rooms). Red square indicates location of the cell processing Facility
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950
949 937
951
952
945
Floor Plan of the CPF9TH floor BRB building
Legend950 – General Area of Development Lab951-Tissue Culture Room952-Tissue Culture Room
Cleanroom Facility937 – Gowning in of the Cleanroom939 – Clean Storage Room
G
946
947
948 938939
941942
944
945-General Area942- Cryogenic Freezer Room944-Gowning out of the Cleanroom946 – Tissue Culture Room947 – Tissue Culture Room948 – Tissue Culture Room949 – Tissue Culture Room
Ancillary Areas938 –Clean Storage Room941 – Cryogenic Freezer storage Room
Products• MSCs – Human
• MSCs – Rat
• MSCs – Porcine
• MSCs – Canine
• c-Kit+ Cardiac Progenitor Stem Cells – Human
• c-Kit+ Cardiac Progenitor Stem Cells – Porcine
• c-Kit+ Cardiac Progenitor Stem Cells – Rat
• c Kit+ Kidney Progenitor Stem Cells Porcine
• Cord Tissue Derived MSCs• Cord Blood Derived MSCs• Placental MSCs• Exosomes• Human and Porcine iPSC• Human T- Cell • Human T-regulatory Cells• Human NK Cells• Amniotic Fluid Stem Cells
• c-Kit+ Kidney Progenitor Stem Cells – Porcine
• Human Dendritic Cells
• Human Tumor Lysate
• Human Schwann Cells
• Rat, Porcine, & Primate Schwann Cells
• gene-modified CD34+ hematopoietic cells
• Human Dental Pulp Stem Cells
Clinical Products
Products being used for clinical trials:
• MSCs
• Bone Marrow Mononuclear Cells
• Schwann Cells
• Dendritic Cells
• Tumor Lysate
• Cord Tissue Derived MSC
• CD34+ Cells
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Mesenchymal Stem Cell Manufacturing
• GLP Manufacturing
• GMP Manufacturing
• Enabled 15 phase I/II INDs
Manufacturing Steps of MSCs
Bone marrow aspira on
Bone marrow
Ini al seeding
Ficoll gradient
MNCs
Expansion
MSCs
CD105: 99.9%
Back to
human
Manufacturing Steps of Schwann Cells • GLP
Manufacturing
• GMP Manufacturing
• Enabled 3 phase I/II INDs
Dendritic Cell Vaccine for Malignant Gliomaand Glioblastoma, sarcoma and brain tumor
• GMP Manufacturing
• Enabled 3 phase I/II INDs
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MULTICENTER CLINCAL TRIALSCardiovascular Cell Therapy Research Network (CCTRN)
Endomyocardial BiopsyAutologousAutologous
C-Kit Cardiac Stem Cells (CSCs)
Digestion ExpansionMagnetic Selection
cGMP Compliant ManufacturingcGMP Compliant Manufacturing
CC--Kit CSCsKit CSCs
Safe Safe PotentPotent
CharacterizedCharacterizedFull DoseFull Dose
• GLP Manufacturing
• GMP Manufacturing
• 1phase I/II INDs at ISCI
• Manufactures for 7 center for phase II trial
Examples of iPSC studies at ISCI
Basic Research (Human and mouse iPSCs)
Preclinical Research(Human and swine iPSCs)
• Modeling molecular and cellular • Development of clinical grade• Modeling molecular and cellular mechanisms of myocardial lineage development
• Development of clinical-grade, autologous and allogeneic iPSC-derived cardiomyoblasts for cell-based therapy in response to heart damage
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Clinical-grade manufacturing of iPSC-derived CPCs expressing the novel receptor
Human and porcine iPSC MCBs from cGMP-grade
MSCs Establishment of xeno-free cGMP
compliant monolayer differentiation
protocols
Scale-up differentiation
and purificationof CPCs
Quantum System (Terumo)
SOP and quality control Porcine model of MISOP and quality control documents
Safety, identity, purity, viability
Assay Qualification
Xeno-free, Virus-freeFeeder-free, Transgene
Free MCB
SOP and quality control documents
Safety, identity, purity, viability
Assay qualification & Validation
Xeno-free, Virus-freeFeeder-free, Transgene
Free product
Development Capabilities
Lab Capabilities
IncuCyte® applications for cell monitoring & workflows
Cells are monitored non-invasively, within the incubator and images are automatically analyzed so investigator can always see what happened when and where.
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Quantum® Cell Expansion
System
QiIACube
This instrument allows for high-throughput extraction of DNA and RNA.
Automated cell culture platform that can help simplify the open, labor-intensive manual tasks associated with flask-based culture.
MoFlow Cell Sorter High-speed sorter, compatible with array of laser options. High viability, yield and purity with an analysis rate of 100,000 events per second and sort rates of up-to 70,000 events per second are standard.
Thank you
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The Center for Biomedicine and Genetics at COH
Joseph Gold, Ph.D.Director, manufacturing
March 14, 2017
Center for Applied TechnologyBeckman Research Institute, City of Hope
GMP Capacity at COHCenter for Biomedicine and Genetics (CBG)
•Designated NGVL Sept, 2001
•Designated ICRC, Sept, 2001•Designated PACT May 2010; Aug 2016•CIRM TC partner facility October 2017
2000
C ll h d i C (C C)
20,000 ft2
GMP Manufacturing Core at COH
2012
•Small molecules
•Reference standards
•Oligo therapeutics
3,000 ft2
2010
Cell Therapy Production Center (CTPC)
•Cadaveric islet cells
•CAR‐T work6,000ft2
Chemical GMP Synthesis Facility (CGSF)
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CBG production experience
• 17 years in operation
• >400 released product lots
Lentivirus
Neural stem cells
hiPSC
hESC
Islet cells
MVA
AdenovirushESC/hiPSC ‐cardiomyocytes
hESC‐RPE/NSC/DA neurons
Small molecule fills
Biologics fills
MSC
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Notable product expertise and capabilities at CBG
• Lentivirus production 100L scale High transduction efficiency
• Neural stem cell modification and banking
• hPSC culture and banking
• hPSC differentiated products: production and dose prepsp p p p
Scalable suspension adapted cardiomyocytes NSC, NPC, DA neuron progenitors Retinal pigment epithelia
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hESC on CELLstart(or Matrigel, Synthemax)
single cells
Accutase
125-1000 mL spinner flask
ROCK inhibitor
StemPro SFM or E8
(40ng/mL bFGF)70 rpm
passage cells every 3 days
Scalable hPSC suspension culture
MCB/WCB
2.5x105
cells/ml
(Accutase)
200 mm
cell aggregates
Vincent C. Chen et al., Stem Cell Research, May, 2012
(Accutase)
differentiation process
•Defined reagents
•No substrates
•No beads, matrices, gels
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d5
cell harvesting
d0 d1 d3 d18-21d2 d4 d6 d7d-2
hPSC expansion
StemPro hESC SFM
cardiac differentiation
RPMI + B27-insulin RPMI + B27
IWP4 CHIR 99021
Wave bag cardiac differentiation
d20(4X)d0 (4X)
Vessel cells/L input cells/L output cell viability cTnT hPSC‐CM/LCardiac/hESC
2L bag 2.5E+08 1.78E+09 96.1% 92.9% 1.65E+09 6.6
cTnT TE7 OCT4
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Target: Macular degeneration (CPCB)Strategy: hESC‐RPE monolayer on synthetic substrate
hESC‐RPE
Parylene substrate
Day 9 D103
hESCMCB
ExpandEnrichDifferentiate ICB
Cryopreserve
hESC‐
RPE
hESC‐
RPE
hESC‐
RPE
Thaw/culture on Parylene
7 Months 1 Month
Full testing
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Wolfram ZimmermannDeepak Srivastava
Academic Collaborators and Clients
Kevin Healy
Tim Nelson
Joseph WuCharles Murry
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Mark Humayun
Production Assistance for Cell Therapy CIRM Translating Center
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Moffitt Cancer CenterCell Processing Facility
Linda L. Kelley, PhDy
Director
Moffitt CPF Staff and Facility
Project ManagementMoffitt Cell Therapies
Cheryl Cox, BSManager
Emily Hopewell, PhDAssistant Technical Director
Nermin Gerges, PhDDevelopment Specialist
Renee Smilee, BSFacility Operations Manager
Martha Hackett, MSQuality Assurance Director
Virginia Garcia-Gobaira, MATraining Specialist
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T Regulatory (Treg) Cells for GVHD
Dendritic Cell (DC) Vaccines
Tumor Infiltrating Lymphocytes (TIL)
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Center for Cell & Gene Therapy cGMP Activities
Adrian GeeBaylor College of Medicine
Houston, Texas
Location
Feigin Center, TCH
Support of Blood & Marrow Transplant Programs
30 beds expanding to 60
• Bone Marrow
• PBPC
• Cord BloodCord Blood
• DLI
• CD34 selected
Special Protocols
Bellicum
Novartis etc.Jeannette Bloom et al.
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Virus‐specific T cells• Used to prevent or treat viral infections inimmune‐suppressed patients
Cliona Rooney
Specialized Products & Services
patients• EBV, followed by CMV,Adeno, HHV‐6, BK & HIVvirus
• Used across HLA barriers
• Manufacturing time reduced from 3 months to 14 days
HelenHeslop
Ann Leen
Chimeric Antigen Receptor (CAR) T Cells
• Directed towards solidtumors
StephenGottschalk
Specialized Products & Services
tumors
• Overcoming tumor resistance mechanism
• Combining tumor and viral specificity to prolong persistence &provide means to reactivate
Maksim Mamomkin
NabilAhmed
Mesenchymal Stromal Cells ‐MSC
• Regenerative and Immunosuppressive
Zhuyong Mei
Specialized Products & Services
Immunosuppressive actions in
• Cardiac disease
• Stroke
• Parkinson’s Disease
• Kidney Transplantation
Adrian Gee
Mariola Klis
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Cell Culture Systems• Simpler and automated systems formanufacturing of
JuanVera
Specialized Products & Services
manufacturing of therapeutic cells
• G‐Rex bioreactor
• Gather
Suicide Genes• Designed to destroy cells in the case of anadverse reaction
MalcolmBrenner
Specialized Products & Services
adverse reaction
• Activated by administration of a small molecule drug
• Used to switch off tumor‐directed T cellsin cases where they may cause GvHD
Therapeutic Cell Sorting• Sorting of Aldehyde‐bright progenitor cells April Durett
Specialized Products & Services
• Used by the CCTRN to treat peripheral arterydisease
• PACE protocol – some promising results
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Quality Control • Endotoxin
• MycoplasmaDeborah Lyon
Specialized Products & Services
• Bioburden
• Replication‐competent virus detection
• Viral titers
• Vector identity
• Stability testing
• EBV assays
Specialized Products & Services
Quality Assurance Services
• Generation of SOPs and worksheets for
Sara Richman
specialized protocols
• Review of records
• Generation ofCertificates of Analysis
• Assistance with CMCsection of IND
NatashaLapteva
MygiaoLe
Contact us at: [email protected]
through the PACT Program
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APPLICATION PROCESSAPPLICATION PROCESS
Debbie WoodEmmes Corporation Rockville, Maryland
PACT Application ProcessPACT Application Process
Web‐based Application• Scope Review by NHLBI
• Full Application invited if scope is approved
Scientific Review Board Review & NHLBI Vote
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TO‐RFP issued; competitive bids
TO awarded (1 CPF)
Contract, Timeline & Budget
Scope Review Application ContentScope Review Application Content
Investigator information (Biosketch, CV, etc.)
Scope
• Relevance to NHLBI scientific interests and PACT Scope
Proposal Overview
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Proposal Overview
• Synopsis of proposed study
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Full Application ContentFull Application Content
Investigator information (Biosketch, CV, etc.)
Scope ‐ Scientific interest and programmatic scope
Proposal Overview
Product information
Current funding ‐ product manufacturing and other activities/tasks not covered
Scope Review
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g p f g /by PACT.
Manufacturing information
Production methodology
Product development
Regulatory information
Timeline
Estimated budget
Institutional/Organizational business office information
Application Review/CriteriaApplication Review/Criteria
NHLBI and an external, independent Scientific Review Board (SRB) comprised of cell therapy experts review:
• Relevance to the scientific mission and programmatic scope
• Applicant and the collaborative team qualifications
• Evaluation of the strengths and weaknesses of preliminary data,
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manufacturing feasibility, significance to the field, degree of innovation,and overall scientific merit of the application
• Detailed translational development plan summary
• Product specifications (proposed release criteria and potency assay)
• Supplemental manufacturing funding
• Demonstration of an adequate plan for eventual use of the cell therapyproduct in a clinical trial
• Proposed work supports regulatory pathway for this product
Next StepsNext Steps
Contract
• Contract negotiations with designated facility
• Intellectual property and/or indemnification
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Timeline
• Development of production task order
• Scheduling of manufacturing tasks
• Scheduling of production
Budget
• CPF assistance
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Website Website
www.pactgroup.net
• Online application process and information
• Education Initiatives
/ d
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• Events/Updates
• Links to PACT facilities, regulatory references & other resources
Contact InformationContact Information
Debbie Wood or Lani IbenanaPhone: 301‐251‐1161Email: [email protected]
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