Be Like Gretzky - Part 2

Part 2

“I never skated to the puck. I skated to where the puck was going to be.”

Quote attributed to Wayne Gretzky ?

In Part 1 I started with this:?

???????????I love that quote.?And I’ll have to admit that, in the past, I’ve over-used that quote.?And I love it whether Wayne Gretzky really said it or not. (It reminds me of Yogi Berra, famous for his convoluted quotes that somehow ring brilliantly true: “I didn’t really say all those things I said.”)??In the article that is the source of the photograph, the author laments that the over-use of the quote by CEOs made it into a trite cliché. But the quote carries a lesson: you must always look ahead in a rapidly changing field.?

Let’s Continue

???????????I wrote a previous article that spring-boarded off that quote to argue that anyone in biotech must not only keep up-to-date with developments in the field, but must be on the look-out for the next big thing, the next innovation.?And the ability to see the future is a special requirement for academics who are setting up training programs.?The academic curriculum review and program approval process often takes a fair amount of time.?Therefore you have to start designing the program a few years, or even years, before the industry matures and needs your graduates as employees.?It involves keeping an ear to the ground, and perhaps a bit of clairvoyance, to design a training program that fulfills workforce requirements a few years out – or perhaps many years out.?Sometimes you just have to guess.?I’ve been lucky that for the most part my guesses, and that of my community college biotechnology professor colleagues, have been correct.

???????????I wrote before that my first guess (and this was heavily influenced by consulting with people from our partner Genentech) was that focusing training on the knowledge and skills required for biomanufacturing was the appropriate emphasis, rather than running an education program that focused on biotechnology research techniques.?This turned out to be right.?Then we developed a program that emphasized single use technologies – and that turn out to be right.?And then we jumped into cell and gene therapy – and that turned out to be right.?And when we designed, with MiraCosta College, the Bachelors of Science in Biomanufacturing program we incorporated Quality by Design, Design of Experiments, engineering principles that emphasized bioprocess monitoring and control – that turned out to be spot on.?With the addition of Total Quality Management Principles, Six Sigma and Lean Manufacturing, and Program Management, and discussions of Enterprise Resource Planning – yeah, those guesses were good too.

???????????When I go to conferences (and I’m often the only non-industry academic in the room), I sit back and think – we nailed this.?

???????????For years we have made the point of emphasizing to students that the products of biotechnology are proteins.?Even though lectures often focused on DNA and recombinant DNA, that bit of genetic engineering was only useful if it could lead to the production of a pharmaceutical protein that could elicit a physiological effect once injected.?

Or, as author and national biotechnology education thought leader Ellyn Daugherty always said:?

DNA is the flash, protein is the cash

DNA is the show, protein makes the dough

???????????Students would often get caught up in the intellectual interest of genetically engineering the cell – the DNA – and would de-emphasize the downstream processing of the protein – the product that makes the dough.?

???????????This paradigm is changed.?In alt protein that product is still protein, but the protein there is for nutrition and eating rather than for being injected as a medicine.?In plasmid production the product, which will serve as a starting material for some other application, IS DNA – in that case the DNA does make the dough.?In cell therapy the product is cells.?In gene therapy the product is a virus that carries the transgene that when injected will compensate for the gene that is damaged or missing in the inherited genetic disorder.?In cultivated meats the product is muscle or fat cells.?And many other examples of protein and non-protein commodity products are about to burst upon the scene – fibers, chemical raw materials currently being extracted from petrochemicals, nutriceuticals, and fuel.?The current number being quoted forecasts a $30 trillion bioeconomy, and that’s a really big number.?

Alternative Protein

???????????In this emerging field of alt protein, the protein remains “the cash”.?Traditional dietary protein is expensive and often environmentally destructive to produce.?Every pound of most meats demands an extraordinary amount of water and energy to produce.?An extraordinary percentage of the land mass of the United States, and of other countries, is devoted to the production of meat.?Alternative proteins hold the prospect of reducing the input and impact required to produce this important component of diet. ???

This field was kicked off by the wildly successful Impossible Foods.?I often choose an Impossible Whopper at Burger King – I can’t remember the last time that I’ve had a beef Whopper, but it’s been a while.?These burgers are anchored by plant product but have been supplemented with a plant form of hemoglobin called leghemoglobin that has been produced by genetically engineered yeast – the inclusion of this protein makes the burger seem more like its animal derived counterpart.?Impossible Foods has also partnered with Starbucks to have that chain carry the Impossible Breakfast Sandwich.?Partnering with successful (and giant) chains makes business sense to increase consumer awareness and the impact of the product.?

???????????One of my favorite companies is Perfect Day Foods.?Perfect Day uses genetically engineered filamentous fungi to produce milk proteins. They produce ice cream and sour cream without cows.?

???????????The company Turtle Tree is similar.?They produce particular milk proteins, like lactoferrin, that currently are not in baby formula.?As it turns out most milk proteins not only serve as a protein source of amino acids, but each protein also has an alternate function – they are a two-fer.?The lactoferrin protein made by Turtle Tree is a good example – it carries iron ions as well as delivering amino acid nutrition.?

Joywell Foods goals is to produce the sweetening protein from miracle berries as a low calorie artificial sweetener.?I have tried yogurt sweetened with it and it’s good.?

The Better Meat company produces a fungal protein that they call rhiza.???According to them “Rhiza mycoprotein contains more protein than eggs, more iron than beef, more fiber than oats, more potassium than bananas, and naturally contains vitamin B12.”?When they talk about it, they refer to Rhiza affectionately, and call her by name, as if she’s a familiar friend.??

???????????Many of the cultivated meat and alt protein companies say that they aren’t really a food company, they are a “fighting global warming” company.?As an example:?“This is why I think people are increasingly aware plant-based products are going to completely replace the animal-based products in the food world within the next 15 years. That’s our mission. That transformation is inevitable,” said Pat Brown, the CEO of Impossible Foods.?

Alternative Fat

???????????We have been sending graduates to an innovative company, Mission Barns, based in San Francisco.?Most everyone that I’ve met there is young and passionate about providing “meat, without the animal.”?They provide the fat cells that make plant-based meat substitutes more savory and lifelike.?They have Cultivated Bacon – plant protein and cultivated pork, Cultivated Meatballs, Cultivated Sausage, and Cultivated Chorizo. “For the first time in history we can cultivate meat without harm.”?When I visited their plant they told me, “you can eat meat where the animal is still alive.?Their webpage states that “Conventional meat production is a major threat to humanity and to our planet,” and everyone that I met there is committed.???

Cultivated Meats

For most of the history of the Solano College biotechnology program, graduates when into biopharmaceutical manufacturing.?We’ve sent hundreds of graduates to Genentech, and hundreds of graduates to BioMarin, and then some to other companies.?Now fully one half of the graduates of the Solano College program are going into cultivated meats.?The field still seems to be struggling for its name.?Laboratory meats has been rejected.?There’s an annual conference called “Cultured Meats and Future Food.” And I’ve heard “cellular agriculture.”

Neighboring UC Davis has emerged as a leader in this field (along with Tufts and UCLA).?They received a multi-million grant to promote the field.?And they use “Cultivated Meats and Alt Protein,” so that’s what I’ll go with.?

The approach used by Mission Barns I discussed above.?We are also sending quite a few graduates to Upside Meats, a company in Emeryville dedicated to growing chicken cells – without chickens - in bioreactors. I was counseled early in my career to look to the people that a company hires to gauge whether that start-up will emerge as a leader; Upside has hired some of a company called Cultured Decadence that has dedicated itself to growing seafood, especially lobster.?You would think that the crab fishery collapse would make the pursuit of that a viable business target.?And this symbolizes why this pursuit is a good idea – conventional agriculture and natural resource harvesting techniques cannot really sustainably supply the demand for meat.????????

Finless Foods generates tuna suitable for sushi without going to the ocean and depleting wild stocks of fish.?Most of the fisheries of the world are endangered, and it isn’t clear that the fisheries would recover with fishing just stopped.?

These companies grow animal cells in giant bioreactors, just like we’ve been doing with Chinese Hamster Ovary cells since the 1980s to produce protein biopharmaceuticals.?The challenges are the same, but different.?Cost is key.?

The key to the entire enterprise is controlling the costs of production; while consumers might give a small premium, the cost must be competitive with meat from animal sources.?The nutrient components of the media, the solution surrounding the cells that provides energy and raw materials, cannot be the lab reagent grade chemicals used at the laboratory scale or used in pharmaceutical biomanufacturing.?In pharma, compliance with regulatory requirements is primary and cost is secondary.?Cultivated meat production will have to use food grade ingredients to save cost because the final cost of the product will be the primary considerations.?The costs aren’t just the components providing nutrition to the cells, but also the protein growth factors that interact with the cells and allow them to divide.?Without those components, and they are currently very expensive.?Every part of the process will have to be optimized.?The cost must be competitive with meat from a butcher.?And that meat’s cost will continue to increase, especially if governmental subsidies are eliminated.?????

???????????The ability to generate meat generated from cells from animals that may still be alive allows palette choices only limited by imagination.?Would anyone like tiger tacos or panda patties??How about lion lasagna??Manta meatballs? Rhino pastrami? I’d like to order a hyena burger? Cultivated caviar?

???????????The control that this production allows could eliminate the risks from many of the food-borne diseases

Plasmids – DNA as a product

???????????Again, for years our key saying has been:

DNA is the flash, but protein makes the cash.?

Rules are made to be broken, and this rule is no exception.?With the production of plasmids, the DNA makes the cash.

Plasmids are small circular rings of DNA replicated in the interior of bacterial cells; they replicate independently from the bacterial chromosome, the main DNA of the cell that codes for all of their proteins.?Plasmids are typically 1/1000 the size of the bacterial genome.?Recently there have been many clever commercial applications for these pieces of DNA. ??

???????????The most successful recent application that used these was the production of the Moderna or the Pfizer/BioNTech RNA vaccines.?These companies grow E. coli cells with plasmids containing the gene that codes for the coronavirus spike protein, the main antigen that generates the immune response.?They grow the bacteria in large tanks, then break the cells open, isolate the plasmid DNA in a pure form, and then place that DNA into a tank with enzymes that read the DNA and use it to make RNA complementary to the spike protein gene.?The RNA is then wrapped in lipid nanoparticles and frozen.?For the generation of these vaccines, plasmid DNA served as the starting material.??

???????????Several plasmids, three to four types, are used to produce the viruses used in both cell and gene therapies. Lentiviruses must be generated for cell therapy; for immunotherapies these viruses carry the gene into the cell that codes for the chimeric antigen receptor that will be embedded in the cell membrane and will enable the killer cell to recognize the cancer cell.?Plasmids are introduced into either insect cells or human HEK293 and give these production cells the ability to manufacture all of the viral components and assemble them into an adeno- associated virus for use in gene therapy applications.?The virus is the product that carries a missing gene into the cells of a patient with an inherited genetic disorder.?

???????????Sutro Pharma has an interesting system to making protein pharmaceuticals – they make proteins without cells.?The use plasmids containing a gene and mix it in a tank with all of the cellular components that enable the gene to be transcribed and the RNA to be translated.?This in vitro approach allows more control and unique design features including the introduction of novel amino acids.?

???????????With all of these applications, in vitro transcription for RNA production (and there are many potential therapeutic strategies besides RNA vaccines), plasmids for the production of viruses, or cell free protein production, the need for plasmid manufacturing facilities is increasing.?I’ve heard at conferences that there is a shortage of manufacturing facilities and increased opportunities for Contract Manufacturing Organizations.

Exosomes

I’m not sure that I had ever heard the term exosome when I was a graduate student.?These small membrane enclosed spaces can carry therapeutic proteins or other molecules.?The Gardasil vaccine against human papilloma virus uses exosomes that had been produced in yeast cells.?

There are multiple companies that are growing mammalian cells and collecting the exosomes produced by the cells.?

Cells or Viruses as a Product

???????????Part 1 of this series discussed producing viruses a gene therapies to deliver functional genes into tissues; viruses are made of multiple proteins are a order of magnitude more complicated than the protein pharmaceuticals that have dominated the field until now.?Cell therapies like CAR-T therapies, where the product is a cell, are orders of magnitude more complicated than that.?Cultivated meats use cells as the product.?And stem cells are about to emerge as a dominant treatment for particular conditions.?

???????????The skills required to work in any of these fields became well established since the 1980s and the production of proteins using mammalian cells, mainly Chinese Hamster Ovary cells.?Workers must learn an array of skills and behaviors collectively called “sterile technique” or “aseptic technique.”?All equipment must be sterilized – and there is a formal definition – there is no living thing, none, nil, zip – and the sterilization process was validated – proven through a very formal procedure to process a state of sterility.?Workers that can do this will never be unemployed.?

???????????And then there are stem cells.?But that discussion deserves its own treatment in the next article.??