“Modern Times” for lactoferrin
Lactoferrin has been called a ‘miracle protein’ for its many beneficial activities, that include anti-bacterial and anti-viral, anti-inflammation and anti-cancer, and more, as we will see below. About 20 years ago we were working with human lactoferrin made in recombinant systems. Now, we hear little about those projects, and much about bovine lactoferrin, so we set out to find out what has been happening with those molecules. Here we share what we found out.
Lactoferrin is present in several human tissues and secretions, including blood plasma, tears, and human milk. Animal versions of lactoferrin (homologues) have also been characterized, in particular, lactoferrin present in the milk of various animals.
The perceived potential for medical applications of human lactoferrin meant that several companies and teams had an interest in developing a scalable process to source the protein. As it is usually the case with human proteins, using the native source is not an option, and although lactoferrin purified from human milk can be purchased, scaling its sourcing using human milk is really not a commercial nor a medical option.
To develop lactoferrin as a potential therapeutic molecule, genetic engineering was used to make recombinant versions of human lactoferrin, starting in the 1990s’. These recombinant human lactoferrin molecules were produced in a broad variety of expression platforms, including in yeast, filamentous fungi, transgenic plants and transgenic animals.
About recombinant human Lactoferrin
While recombinant forms of human lactoferrin have been produced in a broad variety of systems, only three commercial processes for recombinant human lactoferrin (rh-LTF) were developed: production in filamentous fungi, by Agennix (USA and Germany), production in the milk of transgenic cows, by the company Pharming B.V. and production in the grain of transgenic rice by Ventria Inc. Other companies and teams have successfully expressed rh-LTF in various other systems, but they have not progressed to commercial scale.
A very considerable body of research showed that lactoferrin can promote growth of specific cells, inhibit cancer cell growth and has anti-microbial activity. In contrast, it has been hard to demonstrate a clear-cut clinical outcome in a trial and despite several studies, no clinical Phase 3 has shown success. As with other ‘miracle’ proteins, the proposed modes of action for lactoferrin likely could benefit from a systems approach, as this protein likely impacts several metabolic and regulatory pathways.
Spanning several years between 1990s’ and 2010s, Agennix conducted clinical trials with their rh-LTF in a range of indications (including renal cell carcinoma, NSCLC, sepsis). Safety of their protein, which was administered orally, or in some trials, formulated in a gel, for topical application, was demonstrated. In 2002, DSM and Agennix allocated a fermentation facility at a DSM’s site in Italy, to scale up production of this rh-LTF “from tens of kilos to tons”. However, none of the trials demonstrated a conclusive effect, and Agennix lactoferrin (talactoferrin) was never approved as a therapeutic. Agennix ceased operations in 2013. To our knowledge, no other company licensed this product and it was not further developed.
Pharming and a group of other Dutch partners received in 2009 a grant from the government of the Netherlands, to develop their recombinant human lactoferrin produced in transgenic cow’s milk as a functional food for people suffering from inflammatory bowel disease. We have found no publication of study initiations or any research outcomes in this connection yet.
Ventria conducted early-stage trials of its rice grain-produced rh-LTF in children and in adults with diarrhea, but due to regulatory matters, the trials were not continued. Ventria’s subsidiary InVitria has been commercializing the use of its rh-LTF for human stem cell culture and for biomaterials research.
An academic team showed that the rice-made rhLTF lead to skin keratinocyte proliferation in a study of the potential role in wound re-epithelialization. Other studies have shown both rh and bovine LTF to have positive effect in wound healing.
PharmaReview is a Texas-based corporation that is working with a CHO version of rh-LTF. A few publications, including recent ones (2022), describe in vitro studies with this protein, but there is limited information about the future plans for its use on the company’s website. We could not find information about the status of manufacturing, clinical nor commercial development of this protein. As they use CHO for production, there will be many options to produce, when the company needs to scale up manufacturing.
There were discussions about developing infant formula containing rh-LTF. Currently, bovine LTF is used for this product and there are plans to include recombinant bovine LTF in the formula. Infant formula is a very particular product, and there are many ethical considerations that need to be addressed when developing it commercially. This discussion is not in focus here.
What about bovine lactoferrin?
The range of recent reviews about lactoferrin, including assessing the molecule in a COVID setting, shows that interest in the “miracle molecule” has not abated. Rather, the prompt availability of lactoferrin purified from cow milk whey or semi-skimmed milk from a range of suppliers, supported a shift in interest from use of the human protein to the cow milk lactoferrin, for applications in nutrition, skin care and various medical indications. Clinicaltrials.gov lists several ongoing trials with bovine LTF. In clinical trials, bovine LTF administered orally or topically, has been shown to improve various dermatosis conditions. Bovine LTF is also used in cosmetics and as a nutrient. The protein has been used as a source of iron in nutrition, as each molecule of lactoferrin can bind 2 iron molecules.
Is an animal-sourced protein going to replace a recombinant human version, in human therapeutics? This is an interesting and unusual turn of events.
In cow milk, LTF, which has 68% amino-acid homology to the human lactoferrin, is present in concentrations ranging from 20-200mg/L, averaging at 100mg/L of milk. Bovine LTF is purified from whey, a by-product of the cheese industry, or from skimmed milk.
While the price of the primary raw material (cow’s milk) is low, bovine LTF represents about 1% of the whey proteins, so its purification process is costly. Furthermore, LTF in cow’s milk is easily denatured by heat sterilization, making it difficult to retain its bioactivity in commercial milk and dairy products. By extracting LTF from dairy materials and sterilizing it under conditions that do not lead to denaturing, it became possible to incorporate bioactive bovine LTF into various products. Given its important properties, purified (95%) bovine LTF reaches an interesting price (more about prices below).
By the end of 2017, seven bovine lactoferrin manufacturers had obtained Novel Food (EU) certification and three manufacturers, two of which are for infant use and one is for adult use, had obtained GRAS status (USA) for their bovine lactoferrin. The specifications of these regulations are relatively high, including achieving more than 95% lactoferrin purity in protein.
Three of the global producers of bovine LTF are:
Morinaga Milk Industry (Japan), one of the oldest and the largest global producer, has been industrially producing bovine lactoferrin in its fully own subsidiary Milei GmbH, Germany, since 1989. In 2020, Morinaga announced an investment of 15 mio Euro in the Milei site, to more than double the existing production capacity. With this expansion, Milei’s capacity is expected to increase to approximately 170 metric tonnes (or about 150 metric tonnes, if they are using non SI units).
Friesland Campina in the Netherlands, holds Novel Food certification for its bovine LTF. The company announced in 2023 the construction of a new facility in NL with a cost of 80 mioEuro. According to this company, demand is growing both in adult (mainly US) and early life (mainly Asia and China). The company observed that bovine LTF uses in oral hygiene, weight loss and skin health are increasing. The new facility will allow Friesland Campina to increase its production scale from 20 metric tonnes to 80 metric tonnes per year.
The Yili Group from China, one of the largest dairy producers in the world, is the owner of Westland Milk Products in New Zealand. The company has invested in 2023 in the construction of a new facility for bovine LTF production in New Zealand. According to the companies’ websites, the new facility, estimated to be finalized in 2024, will significantly increase production capacity at Hokitika, NZ from a current 20 metric tonnes per year (added capacity not revealed).
Enters the precision fermentation-made recombinant bovine LTF
How will the current combined trends of reducing animal-based foods, including dairy products for personal health reasons, and a better general realization of the negative environmental impact of dairy farming impact the market growth of bovine LTF?
It will be interesting to see if the products issued from precision fermentation will fulfill the new demand and eat into the bovine LTF supply.
Who are the newcomers?
Turtle Tree raised its seed finance in 2020, with a series A the following year that amounted to more than 30 million USD. The team funded the company with the idea of using mammary cells in culture to produce milk. The challenges associated with, amongst others, scaling up the process were considerable, which amongst other aspects of the initial business idea, triggered a review and refocus on valuable milk proteins, starting with LTF. Turtle Tree thinks their rb-LTF process allows them to be on par with the 1000 USD/kg lactoferrin currently reached from bovine LTF. (note: Biomilq announced in 2021 that it had successfully farmed human breast milk outside the body. We have not found more recent information about its development.).
Turtle Tree is not alone in this field, other companies are investing in precision fermentation to produce dairy proteins, including LTF. De Novo FoodLabs, South Africa, founded in 2021, and (perhaps) Helaina Inc., NY, founded in 2019 (it is hard to understand from the information on their website, if Helaina want to use the human or the bovine LTF for their infant formula products) also communicated that bovine LTF is part of their portfolio of products in development.
A few other start-ups are making cow milk proteins using precision fermentation, and some of their products are already being included in commercialized foods. Some examples of these companies are Perfect Day, Eden Brew, Remilk. Currently, these companies have not announced plans to make LTF. It is not only small innovative teams that are active in using precision fermentation to make milk proteins. The giants of the industry are also interested, as illustrated by the birth of Vivici, a joint venture between DSM-Firmenich and the dairy cooperative Fonterra, that got the green light from the EU in December 2022, and its (undisclosed) seed fund earlier in 2023. The JV has the mission of commercializing dairy proteins issued from the two company’s considerable R&D and scalable fermentation resources, starting with bovine beta globulin. While there is no information on when a LTF will be produced, it is highly likely that this protein is on their portfolio.
How will the future market for LTF look like?
The market for bovine lactoferrin has taken off in recent years: from 45 metric tonnes in 2001 to 185 mt in 2012 and was forecast to grow to 262 mt by 2017 mainly driven by demand for infant formula, coming in particular from China. In 2016, however, there was oversupply, leading to significant lowering of prices. Since then, the demand for lactoferrin has increased and it is projected to continue to grow over the coming years. Producers of cow milk-derived LTF have, as we saw, responded by adding significant capacity to their existing operations. Newcomers are challenging the status-quo with novel products made using technologies that are not animal based. The production scales discussed in precision fermentation are impressive: “Once we get to commercial scale for lactoferrin, we’re talking about 190,000-liter fermentation capacity,” (Turtle Tree), but with no indication of yields, it is hard to evaluate how will that production impact the market.
The current discussions about the future of farmed animals are very passionate. The climate impact of the dairy industry cannot be dismissed, neither can the consumer interest in proteins that do not have animal origin nor the important cultural aspects of rearing animals.
Given the health impact of lactoferrin, it is likely that the use of the (animal) protein will continue to increase, regardless, perhaps, of its origin. On the other hand, demand for rh LTF all but disappeared, except in some academic settings. Perhaps the rb-LTF can replace the human form in therapeutic indications, or instead, maybe initial results with the bovine LTF will re-ignite interest in using the human protein in some specific indications. The future of the lactoferrin market is interesting.