Continuous Manufacturing
MedScope has successfully developed, obtained FDA approval, and launched cGMP commercial products manufactured continuously with feedback control analytics (PAT). Moreover, MedScope has developed integrated optical analytics providing real time downstream continuous measurements based upon multi-variate math models embedded into real time firm ware for rapid response. This was first implemented through FDA Emerging Technology guidance documents in which the analytics were implemented as a warning watchdog enabling at line operators to respond to continuous manufacturing feedback. Furtheer still, MedScope has successfully created off-line remote data view software implemented through OPC and the internet. The result of this continuous manfufacture technique is substantially improved manufaturing throughtput and yield, and highly enhanced product quality in which the entire continuous batch was quality controlled. Last, by manufacturing continuously, process dimension is substantially scaled down (while overall batch size is scaled up by time instead of dimension) thereby significantly improving the consistency of thermal, mass, and momentum transport and thus product yield and quality. MedScope is a rapid development organization, and in most instances can initiate value-based discussions within 24 hours. Please contact us at 802-236-8650 or info@medscope.com.

Continuous Manufacturing Improves Quality and Yield In Lentiviral Vector Manufacture by Increasing Surface to VolumeRatio, Thereby Significantly Improving Lentiviral Transduction Efficiency Compared to Traditional Well Plate Transduction Protocols.
Lentiviral vectors (LVs) provide an attractive method for cell transduction, the process of inserting foreign deoxyribonucleic acid (DNA) into a cell with a viral vector, due to their ability to infect non-dividing cells with stable integration of DNA. However, progress in the field has been severely hampered by low transduction efficiencies of current platforms despite various efforts to improve virus-cell interactions. The multiplicity of infection (MOI) is a commonly used parameter for viral transductions to determine the theoretical number of viral particles that will enter a target group of cells. However, much higher MOIs, and therefore greater quantities of virus, are often used clinically to achieve any significant transduction efficiency depending on the cell type. Previous studies have shown that viral transduction is a diffusion-limited process governed by Brownian motion. With currently utilized methods, due to the short half-life of these self-inactivating viruses, many of the viral particles will never reach the target cells before decaying, resulting in the observed low rates of gene transfer. These barriers are mildly circumvented by using high concentrations of LV, which is both cost-prohibitive and potentially toxic to the target cells. Thus, there is a clear need to develop a more efficient platform for lentiviral transduction to limit the amount of virus necessary to achieve therapeutic levels of transduction. Continuous manufacture can be designed to have a high surface area to volume ratio that allows for small sample volumes to be used. Combined with the ability to perfuse fluids with tight control of flow conditions, continuous manufacture provides a novel technical capability in the gene therapy field that enables cells to be exposed to high vol./vol. concentrations of LV without increasing the amount of virus used, thereby overcoming the diffusion limitations and waste associated with current methods of viral transductions. 

Assessment of Transduction Efficiency To Assess Continuous Manufacturing Process Equipment Design
By using GFP encoding lentiviral vector to transduce non-adherent HSC, flow cytometry can be used to evaluate transduction efficiency as a function of process equipment design during the continuous manufacturing process.

Assessment of Transduction Efficiency To Assess Continuous Manufacturing Critical Process Parameters (CPP)
By using GFP encoding lentiviral vector to transduce non-adherent HSC, flow cytometry can be used to evaluate transduction efficiency as a function of various process critical process parameters during the continuous manufacturing process.

Testing Impact of Continuous Processing Equipment Design and CPP "at-lab" and "in-mice" - CAR-T Cell Therapy Example
Primary human T cells have been transduced in continuous manufacturing process using clinical grade LV encoding for GFP (or a high titer coagulation factor VIII (fVIII) LV.) Transduction efficiency and cell viability were then assessed by measuring GFP expression and 7AAD dead cell staining via flow cytometry. Vector copy number (VCN) was measured by the real time polymerase chain reaction (RT-PCR) to determine true virus integration and efficiency of transduction. Continuous manufacturing transduction efficacy was also assessed in vivo using primary murine Sca1+ hematopoietic stem and progenitor cells from C57BL/6J hemophilia A mice. The isolated cells were transduced in a standard 6-well and in the continous manufacturing transduction system with the fVIII-LV for direct comparison. Transduced cells were then transplanted into lethally irradiated hemophilia A donor mice. Transduction efficiency and engraftment were assessed by measuring fVIII plasma levels via chromogenic assay and flow cytometric analysis of recipient versus donor cells, respectively. Vector copy number was measured from cells of the blood, spleen, and bone marrow after at least 8 weeks when fVIII levels typically stabilized. 

Modification of Existing Cell Culture Bags To Increase Cell Induction (GT or CART)
​By hot embossing patterns into existing cell culture bags, surface area to volume can be greatly increased in continuous manufacturing, thereby enabling higher transduction efficiencies by enabling higher surface area to volume ratio to increase induction rates by exposing more cells to the LV, AND reducing transduction times which reduces cell degradation. Additionally, gas permeable sheets of similar materials to cell culture bags could be used with a xurographic manufacturing process to pattern channels and bond two layers together to form a channel in a bag. 

Characterizing Primary T cell Transduction - Impact From Continuous Manufacturing
Assessment of vector integration as a primary output due to limitations in LV availability is useful in characterizing the TCell transduction process. Additionally useful is transducing T cells with CAR-LV so that functional assays can be conducted for efficacy or safety profiling. With T cells, tumor cell-specific cytotoxicity can be measured to quantify the anti-cancer potential of continuous flow-generated CAR T cells. 

Characterizing Hematopoietic Stem Cell Transduction - Impact From Continuous Manufacturing
Assessment of vector integration as a primary output due to limitations in LV availability is useful in characterizing the HSC transduction process. Additionally useful investigating the efficacy of continuous flow in transducing human CD34+  cells, which are an even more difficult target for genetic modification. The cells could then be assessed with the human colony forming cell assay using methylcellulose-based media to determine if engraftment has been compromised. Alternatively, transduced human CD34+ cells could be transplanted into humanized NOD SCID mice (immunodeficient mice) to assess engraftment potential. Preclinical development of a CD34+ fVIII-LV gene therapy product candidates have been reported to have an order of magnitude reduction in vector costs (due to increased efficiency of transduction) per patient using small scale continuous manufacturing.
















    MedScope - BioProcess Consultants
        QbD/CMC/cGMP Consultants in Gene Therapy, Immunotherapy, Small Molecule, LentiVector Development: Inception to Launch
         802-236-8650     info@medscope.com

Process - Operations
 - LentiVector  -
 - Development/Mnftr
 - Cell Line Development/Mnftr
 - Continuous Manufacturing
 - Process Analytics (PAT)
 - DOE, Multivariate Design
 - Scale-Up,-Out,-Down
 - Development Protocols
 - Equipment
 - Single Use
 - Strategic De-risk

Regulatory Science
 - CMC Development
 - Comparability Studies
 - Device Design Controls

Analytical Development
 - Bioanalytic Assays  -
 - Immuno/PK
 - Product/Impurity
 - Single Use
 - Extractables/Leachables

Medical
 - CMO/Med Affairs
 - Preclinical Development
 - Clinical Development
 - CMC/Commercial 
 - Strategic Develop
 - Rare Disease
 - Pediatric 
 - Breakthrough Therapies

Manufacturing and ComOps
 - Capital Equipment
 - BSL-2, ISO7
 - Closed Manufacturing
 - Suppliers
 - Tech Transfer

Facilities & Locations
 - 100,00 ft2 Mnftr//Lab Space
 - 50,000 ft2 Office Space 
- Cambridge, MA
 - Burlington, VT

Quality
 - Documentation
 - WIFI Controls 
 - QBD, SOP
 - WI, BPR, Protocols, 
 - Audits, PAI, 

Sales & Marketing
 - Global Strategic Pricing
 - General Management
 - Strategic Plan
 - Market Plans
 - Aliance Management

Political Advocacy, Gene Therapy
 - Gene Therapy Cost Models
 - FDA Realignement
 - Workforce Expansion
 - Workforce Development
 - Regs Patient Bedside Mnftr

U.S. Defense
 - Biologics Rapid Tech
 - Monitor/Detect
 - Biologics Rapid Response
 - DARPA, SBIR

Computation
 - Protein/DNA Modelling
 - Process Capability
 - Remote RealTime OPC/OLE










802-236-8650, info@medscope.com