Footprint size matters in cell and gene therapy manufacturing

Cell and gene therapies have demonstrated their extraordinary, curative potential in the last decade. They’ve had the biggest impact in liquid tumors, but with over 1000 clinical trials in the advanced therapy space underway, cures for a range of diseases are on the horizon.

The latest ground breaking news in the space is the European Marketing Authorization for Kite’s Yescarta as a second line treatment for B-cell diffuse lymphoma. Until now, CAR-T therapies have only been approved for refractory patients, with patients having to undergo at least 2 lines of treatment before being eligible. But with the increased focus on getting cell and gene therapies to market, the expansion of approved therapies to wider patient populations, and the use of these therapies earlier in the treatment pathway, manufacturing capabilities need to be drastically improved to meet the oncoming demand.

This opens up a plethora of problems. Aside from the commonly discussed issues; the lack of qualified manufacturing personnel, supply chain issues with reagents and plasticware, the high batch failure rates plaguing manufacturing processes, and the logistics of delivering cell therapies to patients; there simply isn’t enough space for large-scale cell and gene therapy manufacturing. So how big is this problem? Let’s do a rough breakdown:

1 clean room = ~150sqm

à This can run 10 simultaneous batches with the best automated platforms on the market

For a typical CAR-T process with a team working full day and night shifts:

1 batch = 2 weeks

So in 1 year: 10 batches x 2 weeks x 12 months = 240 batches

Currently CAR-T manufacturing represents less than 1% of clean room utilization and the number of patients treated with cell therapies every year is in the thousands. But the addressable population for just CAR-T therapy is going to be ~2 million patients in the next 5-10 years. Which will require:

8,000 clean rooms = 1,200,000 sqm

That’s equivalent to the current total cleanroom capacity worldwide.

In reality, this space should be doubled to factor in space for QC labs, storage space, offices etc, and including the manufacture of non-CAR-T cell therapies this requirement could even be tripled. This substantial need for more space is good news for clean room suppliers. But the space and construction requirements, and the expensive equipment needed to run these clean rooms to regulatory standards isn’t so good for cell and gene therapy developers, patients or the environment.

At MFX we are developing the Cyto Engine™, an automated cell therapy manufacturing platform that can produce up to 100 batches per square meter. That’s more than 30x the number  of batches per square meter than current manual processes, and more than 4x the number of batches per square meter than current automated processes.

How to ensure a consistent cell expansion process? Don’t start at the end

Cell culture has a culture of measuring endpoints. Vessels used to expand cells (like multi-well plates and flasks) are essentially mystery boxes in the days or weeks of culture. It’s only at the point of cell harvesting that most researchers measure and analyse their cells, which means all the information that they need to know – what goes on inside the vessels during expansion, what effect the environment is having on the cells, is the expansion process consistent – remains an enigma.

One of the issues facing sampling during culture is the size of the vessels. For T-25 flasks or multi-well plates, removing enough cells or medium to give accurate readings will disrupt the culture and substantially affect results. For mid-range vessels like large flasks or bags this is less of a problem, but only sampling once or twice per process operation (as is usually the case) doesn’t accurately reflect cell behavior over time.

The next big issue is the analytical equipment needed. Most labs have flow cytometers, cell counters, microscopes, and PCR machines. But few have more advanced equipment like metabolite analyzers, and on-line, real-time measurement capabilities don’t exist for standard research vessels. Sensors that continuously monitor parameters like pH and dissolved oxygen can be found on (very) expensive automated stir tank bioreactors, but when was the last time you saw a T-75 hooked up to a computer?

To understand cell behavior and ensure a consistent expansion process, measuring the parameters that can affect cells during culture is crucial. Existing indicators like color changing medium may provide rough estimations of metabolite build up in a vessel, but oftentimes metabolites accumulate to detrimental levels way before the medium turns yellow, with cell health and growth being hindered by amino-acid deficiencies. This will only become apparent after weeks of tedious pipetting when the cells are analyzed, and if this happens in the same run, researchers will be left none the wiser about why part of their experiment failed.

A solution to this problem is affordable online measurement tools that are intuitive, account for common problems like calibration and drift, and collect data into central, usable formats. But these don’t currently exist.

At MFX we’re developing the Cyto Engine™ – automated cell research and manufacturing platforms with online media and cell measurements. We’re working with the best sensor companies in the industry to integrate their technology and making it easy to account for drift and calibration. Data is aggregated and available from any device at any time, and our partnerships with the best visualization and data mining software providers means the Cyto Engine™ give real, valuable insights into cell and process data and maximize the information that each experiment provides.

New technologies, old methods- Why 99% of cell culture vessels will never scale

“We’re entering a new frontier in medical innovation with the ability to reprogram a patient’s own cells to attack a deadly cancer.”

That’s what Scott Gottlieb, commissioner of the FDA said when the first cell therapy Kymriah® was approved in the US in 2017. Since then, only 23 cell and gene therapy products have been approved by the FDA. Why are so few of these life-changing therapies reaching the market? And when they do, why are they so hard to access? Sure, these new technologies require stringent testing through clinical trials before approval. But one of the biggest issues restricting access to approved therapies- the expensive price tag- is largely due to the inadequate manufacturing methods currently available.

So why can’t existing manufacturing methods scale-up? Let’s look at the development process.

Research typically starts with a multi-well plate. These are good because they’re cheap, don’t consume a lot of reagent and cells, and have a small footprint. But the process is largely manual, which brings in a host of issues from manual handling and makes it hard to monitor what is going on in the wells. And soon, something with a larger surface area is needed to expand the cells in. A T-flask for example.

It may seem obvious to say that a flask has a very different geometry to a multi-well plate, but this change of vessel means cells will be exposed to a different environment. Nutrients, metabolites, paracrine factors, and gasses will all diffuse differently, while seeding, agitation and harvest methods will introduce new shear stress and temperature fluctuations. All these factors will ultimately have an effect on cell density, viability, and phenotype. And now, adjusting the process to redirect cell phenotype towards their original state is going to require more space and reagents.

Once pre-clinical testing is done, it’s time to scale-up for Phase I using a stirred tank bioreactor, culture bag, or even an automated platform that genty rocks cells. All the factors in the previous jump from plates to flasks are multiplied here, so cells are going to look COMPLETELY different. Although there’s more real-time monitoring methods available with bioreactors, even more time, effort, and money will need to be spent optimizing expansion.

“We talked to over 100 cell therapy developers during our development of the Cyto Engine and on average, they will spend 1-2 years translating their process from a research scale (flask) to a manufacturing scale (stirred tank, bag or otherwise)” says Antoine Espinet, CEO & Co-Founder of MicrofluidX, “It really got us thinking –  how many patients could have been treated in that time and how much money is being spent on all this?”

There is huge unmet need for truly scalable cell culture platforms that can translate cell and gene therapies from research to manufacturing scale, keeping consistency of the final product throughout. That’s why we’re developing the Cyto Engine, combining scalable bioprocessing, complete integration of online PATs, and powerful data visualization and analysis. 

99 problems in cell research – manual handling is number one

When you think of a biologist you probably picture someone pipetting in a lab growing organisms out of a Petri dish. Companies have made fortunes selling glassware, plasticware, and the myriad of equipment like incubators, centrifuges, laminar flow hoods, needed to run a cell culture lab. Today, you’ll be hard-pressed to equip a basic biology lab for less than $500k.

But despite modern day lab equipment’s increasingly clean, futuristic lines and connected capabilities, highly skilled individuals remain an indispensable workforce in the lab. This workforce has a high monetary and human cost- biologists need to carefully orchestrate every movement to safeguard sterility while hunched over a BSC for long hours, saying goodbye to their weekends and bank holidays.

Beyond costs and inconvenience, manual handling’s fundamental issue is the variability that it generates. Operators doing the exact same experiment have demonstrated variability in their results, and even the same operator can struggle to get their experiments to repeat. You might be familiar with the old ‘it’s biology- cells are capricious’ argument, but looking at the process you can’t totally blame the cells.

James Kusena, VP of Bioprocessing and Applications at MFX explains; “ Every single manual operation (in cell culture) can add variability; how fast you pipette, where you place the pipette in the dish, the time difference between the first and last well being filled, where things are placed in the incubator, how you agitate the dish, and last but not least, how you measure your data (cell counts, flow panels etc). And that’s before you factor in the occasional (but inevitable) mistake – we are all human after all!”

Might automated cell culture systems be the answer then? Current fully automated systems for process development cost the same as a full lab to buy (and cost an arm and a leg to run). They often lack versatility and user-friendliness, so you end up running most things manually anyway while the automated system sits idle. Well plate automation systems can be useful if you’re not ready to scale, but when you do, you’ll need to prepare for years of pain transferring your process to larger vessels. Investing in a robotic arm helps automate some of the cell culture process, but in the grand scheme of things it’s like inventing a mechanical horse to pull a chariot instead of inventing an automobile.

In our opinion, there is gap in research for a low-volume, fully automated, scalable, and affordable cell culture system.