Ross Peterson

Aging is characterized by progressive dysregulation of interconnected biological systems, including immune function, gut integrity, iron homeostasis, and mitochondrial metabolism, which collectively contribute to inflammaging, oxidative stress, frailty, and age-related disease. Lactoferrin, a multifunctional iron-binding glycoprotein, has emerged as a potential nutritional intervention capable of modulating several hallmarks of biological aging. This narrative review examines current mechanistic and clinical evidence regarding lactoferrin’s role in healthy aging. Evidence from in vitro, preclinical, and human studies suggests that lactoferrin supports gut health, iron regulation, and immune function and may support mitochondrial function and redox balance. Although much of the mechanistic evidence remains preclinical, early human studies demonstrating improvements in inflammatory and iron-related biomarkers support further investigation. Well-designed clinical trials in older adults are needed to determine whether lactoferrin supplementation can meaningfully improve functional outcomes, cardiometabolic health, immune resilience, and markers of biological aging.

The preclinical safety of human-equivalent lactoferrin alpha (heqLFα; effera ^® ), produced by Komagataella phaffii , was evaluated to support its use as a food ingredient in infant formula and products for young children. Genotoxicity was assessed using a bacterial reverse mutation (Ames) assay in five strains of Salmonella typhimurium and Escherichia coli, and an in vitro micronucleus assay in TK6 cells. Both studies followed OECD guidelines and were conducted up to the recommended limit concentrations (5000 µg/plate and 2000 µg/mL, respectively). A 14-day non-GLP juvenile dose range-finding study and a 13-week GLP juvenile rat toxicity study with a 4-week recovery phase were conducted under ICH S11 guidance. Neonatal Sprague-Dawley rats received heqLFα at 0, 1500, 3000, or 5000 mg/kg body weight/day by twice-daily oral gavage from postnatal day 7 to 98. Bovine lactoferrin (bLF) and whey protein at 5000 mg/kg body weight/day served as reference controls. heqLFα was non-mutagenic and non-clastogenic in both genotoxicity assays. In the 13-week juvenile rat study, no heqLFα-related mortality, clinical signs, developmental, neurobehavioral, or immune toxicity effects were observed. Minor, non-adverse renal findings consistent with high protein intake were observed and largely reversed during recovery. Clinical pathology parameters fully resolved during recovery. Toxicokinetic evaluation showed no systemic accumulation. Based on the absence of toxicologically relevant adverse findings, heqLFα is well tolerated at doses up to 5000 mg/kg body weight/day, establishing a no observed adverse effect level (NOAEL) of 5000 mg/kg/day, the highest dose tested, and supporting its safety across intended populations from birth to adulthood.

Human lactoferrin (hLF) is a glycoprotein of commercial interest as a food ingredient for gut health. We conducted an exploratory analysis comparing the effects of Helaina hLF (effera^®), produced by Komagataella phaffii, and bovine LF (bLF) on the adult gut microbiome and fecal metabolites. In a randomized, double-blind, parallel-arm, controlled trial, 66 healthy adults received either high-dose (HD) effera^® (3.4 g/day), low-dose (LD) effera^® (0.34 g/day), or bLF (3.4 g/day) supplementation for 28 days. Fecal samples collected at Days 0 (baseline), 28, 56, and 84 were analyzed for microbial diversity, taxonomic shifts, and volatile fatty acids (VFA). Alpha-diversity remained stable across all groups. Beta-diversity showed no main effect of treatment; however, bLF was associated with significant visit-related shifts, as assessed by weighted UniFrac. At the phylum level, significant changes associated with effera^® were observed, including decreases in Bacillota (LD) and Verrucomicrobiota (HD), and notable genus-level increases in Lachnospira, Paraprevotella, and Faecalibacterium (HD), while bLF was associated with an increase Bacteroidota at the phylum level and with an increase in the genus Roseburia. Both effera^® and bLF were associated with decreases in Blautia and Dorea. VFA analysis revealed that bLF increased absolute concentrations of total short-chain fatty acids (SCFAs) and branched-chain fatty acids (BCFAs); both effera^® treatments produced proportional changes in SCFAs as a percent of total VFA, acetate, and individual BCFAs. In healthy adults, effera^® supplementation promoted a proportional increase in acetate and supported potentially beneficial taxa while maintaining microbial diversity, without disrupting community structure. As a pre-specified exploratory outcome, microbiome and VFA findings should be interpreted as hypothesis-generating. (clinicaltrials.gov: NCT06012669).

Background/Objectives: Human lactoferrin (hLF) is glycoprotein of commercial interest as a food ingredient for gut health. Here, we report an exploratory analysis evaluating the effects of Helaina hLF (effera ^® ), produced by Komagataella phaffii , on the adult gut microbiome and fecal metabolites in comparison to bovine LF (bLF). Methods: In a randomized, double-blind, parallel-arm, controlled trial, 66 healthy adults received either high-dose (HD) effera ^® (3.4 g/day), low-dose (LD) effera ^® (0.34 g/day), or bLF (3.4 g/day) supplementation for 28 days. Fecal samples were collected at baseline (Day 0), Day 28, Day 56, and Day 84 and analyzed for microbial diversity, taxonomic shifts, and volatile fatty acids (VFA). Results: Alpha-diversity remained stable across all groups. Beta-diversity showed no main effect of treatment; however, bLF was associated with significant visit-related shifts, as assessed by weighted UniFrac. At the phylum level, significant changes associated with effera ^® were observed, including decreases in Bacillota (LD) and Verrucomicrobiota (HD), and notable genera increases in Lachnospira , Paraprevotella , and Faecalibacterium (HD), while bLF was associated with an increase in Roseburia . Both effera ^® and bLF were associated with decreases in Blautia and Dorea . VFA analysis revealed that bLF increased absolute total short-chain fatty acids (SCFAs) and branched-chain fatty acids (BCFAs) concentrations, while both effera ^® groups produced proportional changes in SCFAs, individual BCFAs, and acetate. Conclusions: In healthy adults, effera ^® supplementation promoted a proportional increase in acetate and supported potentially beneficial taxa while maintaining microbial diversity, without disrupting community structure. (clinicaltrials.gov: NCT06012669 ).

Oral toxicity and toxicokinetic properties of human lactoferrin (LF) alpha produced in Komagataella Phaffii (effera™) were investigated in adult Sprague-Dawley rats over a 28-day period under good laboratory practice conditions. Main study dosing used groups of 10 rats/sex/dose, and a secondary study evaluating toxicokinetic parameters used 6 rats/sex/dose. The vehicle control group received sodium citrate buffer, test groups received daily doses of 200, 600, and 2000 mg of effera™ per kg body weight, and the comparative control group received 2000 mg bovine LF (bLF)/kg body weight per day. T-cell dependent antibody response against keyhole limpet hemocyanin and immunophenotyping of the spleen were performed as measures of immunotoxicity. Clinical observations, body weight, hematology, coagulation, clinical chemistry, urinalysis, immunotoxicity, gross necropsy, and histopathology were assessed. Toxicokinetic parameters were analyzed as an indication of LF bioavailability, and anti-LF antibody assays were conducted to detect antibodies produced against LF to measure immunogenicity. No treatment related toxicologically significant changes were observed. Based on the absence of toxicologically relevant changes, effera™ is well tolerated in rats at doses up to 2000 mg rhLF/kg/day, an amount ∼400 times that of the estimated daily intake at the 90th percentile proposed for human adult use.

Background: Recombinant human lactoferrin (rhLF) is of commercial interest for support of the immune system and iron homeostasis but is not currently available as a food ingredient. Objective: The objective was to evaluate the immunogenicity/alloimmunization potential of Helaina rhLF (effera™) from K. phaffii over a 28-day period compared to bovine LF (bLF). It was hypothesized that rhLF would have an equal or lower immunogenic potential compared to bLF, which is permitted for use in conventional foods and infant formula in the EU and US. Methods: Study 1 was a randomized, double-blind, parallel arm, controlled trial where 66 healthy adults were randomly allocated to 1 of 3 groups: high-dose rhLF (3.4 g/d), low-dose rhLF (0.34 g/d), or bLF (3.4 g/d) (clinicaltrials.gov: NCT06012669 ). Participants completed a 28-day (Day 28) supplementation period followed by a 28-day washout period (Day 56) and an additional 28-day follow-up period (Day 84). Study 2 was a 12-week observational study with no intervention that enrolled 24 healthy adults. In both studies, fasting blood was obtained on Days 0, 28, 56, and 84 for analysis of anti-bLF and anti-hLF antibody levels as the primary endpoint as well as other secondary safety endpoints including complete blood count, iron biomarkers, and metabolic panel. Results: In Study 1, the change from baseline to Day 56 in serum anti-bLF antibodies in the bLF group (least squares geometric mean and 95% confidence interval for the post/pre ratio: 3.01; 2.08, 4.35) was greater than the changes in serum anti-hLF antibodies in the low-dose rhLF (1.07; 0.77, 1.49; PTM as a food ingredient at an intake level up to 3.4 g/d in healthy adults.

This work presents a thorough characterization of Helaina recombinant human lactoferrin (rhLF, Effera™) expressed in a yeast system at an industrial scale for the first time.

The oral toxicity of recombinant human lactoferrin (Helaina rhLF, Effera™) produced in Komagataella phaffii was investigated in adult Sprague-Dawley rats by once daily oral gavage for 14 consecutive days. The study used groups of 3-6 rats/sex/dose. The vehicle control group received sodium citrate buffer and the test groups received daily doses of 200, 1000 and 2000 mg of rhLF per kg body weight. Bovine LF at 2000 mg/kg body weight per day was used as a comparative control. Clinical observations, body weight, hematology, clinical chemistry, iron parameters, immunophenotyping, and gross examination at necropsy were used as criteria for detecting the effects of treatment in all groups and to inform dose levels for future toxicology studies. Quantitative LF levels were also analyzed as an indication of bioavailability. Overall, administration of Helaina rhLF by once daily oral gavage for 14 days was well tolerated in rats at levels up to 2000 mg/kg/day, or 400x Helaina’s intended commercial use, and indicating that a high dose of 2000 mg/kg/day is appropriate for future definitive toxicology studies.