This work presents a thorough characterization of Helaina recombinant human lactoferrin (rhLF, Effera™) expressed in a yeast system at industrial scale for the first time. Proteomic analysis confirmed that its amino acid sequence is identical to that of native human LF. N-linked glycans were detected at three known glycosylation sites, predominantly oligomannose structures. Helaina rhLF's protein secondary structure was nearly identical to that of human milk lactoferrin (hmLF), and small-angle X-ray scattering and analytical ultracentrifugation confirmed well-folded globular structures in solution closely matching the reported crystalline structure of iron-bound native hmLF. Differential scanning calorimetry revealed two distinct denaturation temperatures consistently mirroring those observed for apo- and holo-hmLF. Overall, Helaina rhLF differed from hmLF only in its N-glycans, while affirming high purity and globular structures closely akin to hmLF.
