Evaluation of the potential food allergy risks of recombinant human lactoferrin expressed inKomagataella phaffii

Abstract

Prior to the introduction of novel food ingredients into the food supply, safety risk assessments are required, and numerous prediction models have been developed and validated to evaluate safety. The allergenic risk potential of Helaina recombinant human lactoferrin (rhLF, Effera™), produced inKomagataella phaffii(K. phaffii) was assessed by literature search, bioinformatics sequence comparisons to known allergens, glycan allergenicity assessment, and a simulated pepsin digestion model. The literature search identified no allergenic risk for Helaina rhLF,K. phaffii, or its glycans. Bioinformatics search strategies showed no significant risk for cross-reactivity or allergenicity between rhLF or the 36 residual host proteins and known human allergens. Helaina rhLF was also rapidly digested in simulated gastric fluid and its digestibility profile was comparable to human milk lactoferrin (hmLF), further demonstrating a low allergenic risk and similarity to the hmLF protein. Collectively, these results demonstrate a low allergenic risk potential of Helaina rhLF and do not indicate the need for further clinical testing or serum IgE binding to evaluate Helaina rhLF for risk of food allergy prior to introduction into the food supply.

Precision Fermentation as an Alternative to Animal Protein, a Review

“…However, the extent and type of N-glycosylation have a profound impact on the therapeutic properties of many proteins [167][168][169]. Studies have successfully demonstrated a glycoengineering strategy in K. phaffii that enables the production of completely humanized and immunologically compatible recombinant lactoferrin, although with considerably lower yields (Table 2) [133,170]. It is well established that glycosylation of human lactoferrin occurs predominantly at three sites: Asn-156, Asn-497, and Asn-642 [167].…”

“…It is well established that glycosylation of human lactoferrin occurs predominantly at three sites: Asn-156, Asn-497, and Asn-642 [167]. The process to produce humanized lactoferrin involves eliminating endogenous yeast glycosylation pathways, such as some glycosyltransferases, followed by the gradual introduction of heterologous glycosylation enzymes, including mannosidases, other glycosyltransferases, and epimerases [168,170]. This approach contributes to increasing the half-life of the glycoprotein in vivo and eliminates allergenic potential, thus enhancing the therapeutic value of yeast-derived glycoproteins [171].…”

Dose range-finding toxicity study in rats with recombinant human lactoferrin produced inKomagataella phaffii

“…The use of hmLF isolated from human milk has not been pursued due to economic, ethical, and sustainability reasons. Approximately one liter of human milk is required to yield 100 mg of hmLF 17 ; not only is this practice costly and wasteful, but milk used for this purpose could be unsuitable for feeding infants. The expression of recombinant proteins provides a method for production at commercial scale with prospective sustainability benefits, and at a lower cost and without ethical concerns.…”

Dose Range-Finding Toxicity Study in Rats With Recombinant Human Lactoferrin Produced in Komagataella phaffii

“…The use of hmLF isolated from human milk has not been pursued due to economic, ethical, and sustainability reasons. Approximately one liter of human milk is required to yield 100 mg of hmLF; 20 not only is this practice costly and wasteful, but milk used for this purpose could be unsuitable for feeding infants. The expression of recombinant proteins provides a method for production at commercial scale with prospective sustainability benefits, and at a lower cost and without ethical concerns.…”