Human Milk Lactoferrin by Precision Fermentation

Project Information

HuMiLAF

Grant agreement ID: 101218593

DOI

10.3030/101218593

EC signature date 16 June 2025

Start date 1 May 2025

End date 30 April 2027

Funded under

• The European Innovation Council (EIC)

Total cost

No data

EU contribution

€ 2 499 999,00

Coordinated by

PFX BIOTECH LDA
Portugal

Periodic Reporting for period 1 - HuMiLAF (Human Milk Lactoferrin by Precision Fermentation)

Reporting period: 2025-05-01 to 2026-04-30

Summary of the context and overall objectives of the project

The HuMiLAF project (Human Milk Lactoferrin by Precision Fermentation) addresses a growing global need for reliable, sustainable, and scalable production of high-value bioactive proteins. Among these, human milk lactoferrin (hLF) is of particular interest due to its multifunctional biological properties, including immune modulation, antimicrobial activity, iron regulation, gut health support, and overall health-promoting effects. These characteristics make hLF a highly attractive ingredient for a range of applications including infant nutrition, medical nutrition, dietary supplements, healthy ageing products, and functional foods.
Today, access to lactoferrin is constrained by significant supply-side challenges. Commercial lactoferrin is predominantly obtained through extraction from bovine milk, a process that depends on dairy production volumes, complex purification procedures, and volatile global supply chains. As demand for lactoferrin continues to increase worldwide, manufacturers face challenges related to ingredient availability, fluctuating prices, supply security, and batch-to-batch variability. These constraints limit market growth and hinder the wider adoption of lactoferrin-containing products across multiple sectors.
The challenge is particularly relevant for infant nutrition, where lactoferrin is recognised as one of the key bioactive proteins naturally present in human milk. Although infant formula has continuously improved over recent decades, important differences remain between formula and breast milk in terms of bioactive composition and functionality. Limited access to human-identical lactoferrin has restricted the development of next-generation nutritional solutions capable of more closely reproducing the biological functions of human milk. At the same time, increasing scientific evidence supporting the health benefits of lactoferrin is driving demand beyond infant nutrition. Medical nutrition products, immune-support formulations, dietary supplements, and healthy ageing solutions increasingly seek access to high-quality lactoferrin as a functional ingredient. Consequently, the need for a scalable and reliable production platform extends far beyond a single market segment and represents a broader challenge for the nutrition and health industries.
Within this context, HuMiLAF aims to establish a novel precision fermentation-based manufacturing platform to produce human milk lactoferrin. The overall objective of the project is to develop and validate a commercially viable process capable of producing hLF at industrial scale with consistent quality, high functionality, and competitive economics. Building on PFx Biotech’s proprietary technology platform, the project encompasses strain engineering, bioprocess optimisation, downstream processing development, scale-up activities, product characterisation, formulation studies, regulatory preparation, and commercial validation.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

During the first 12 months of the HuMiLAF project, activities focused on establishing a robust and reproducible manufacturing platform for human milk lactoferrin (hLF), validating product quality and functionality, and preparing the technology for pre-industrial-scale deployment. Significant progress has been achieved in Work Packages 1 and 2, while scale-up and formulation activities under Work Packages 3 and 4 have been successfully initiated.
The main technical achievement of the reporting period was the successful completion of the 1,500 L scale-up programme and the demonstration of process robustness across upstream and downstream operations. The project achieved reproducible production of human milk lactoferrin at pilot scale, meeting the requirements defined for technology transfer to the next pre-industrial scale.
Extensive studies were conducted to evaluate strain robustness and reproducibility. The production strain consistently demonstrated stable growth and production performance across multiple fermentation runs, confirming the suitability of the expression system for large-scale manufacturing. In parallel, critical bioprocess parameters were systematically evaluated and optimised, including pH, temperature, aeration, overpressure, and feeding strategies. The results demonstrated broad operational robustness and enabled the definition of a well-controlled production window suitable for scale-up.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

HuMiLAF is advancing beyond the state of the art by demonstrating that human milk lactoferrin can be produced through precision fermentation using a process specifically designed for industrial scalability and commercial implementation. The successful scale-up to 1,500 L represents an important validation of process robustness and reproducibility and confirms that production performance can be maintained under industrially relevant operating conditions. This achievement substantially reduces one of the key technical risks that has historically limited the commercialisation of recombinant bioactive proteins.
Beyond scale-up, the project has generated advances in process design and manufacturing performance. The optimisation of upstream and downstream operations has resulted in a highly controlled production process capable of consistently delivering high purity levels and strong recovery yields. These achievements contribute directly to improving manufacturing efficiency and represent important steps towards achieving the production economics required for large-scale market adoption.
The project has also strengthened the scientific understanding and validation of precision fermentation-derived human lactoferrin. The results obtained so far indicate a structural similarity to human milk lactoferrin and clear differentiation from bovine lactoferrin. Functional assessments performed during the project further support the biological relevance of the produced protein and provide an important foundation for future product development and regulatory activities.