Human intrinsic factor expressed in the plant Arabidopsis thaliana

Intrinsic factor (IF) is the gastric protein that promotes the intestinal uptake of vitamin B-12. Gastric IF from animal sources is used in diagnostic tests and in vitamin pills. However, administration of animal IF to humans becomes disadvantageous because of possible pathogenic transmission and contamination by other B-12 binders. We tested the use of recombinant plants for large-scale production of pathogen-free human recombinant IF. Human IF was successfully expressed in the recombinant plant Arabidopsis thaliana . Extract from fresh plants possessed high B-12 -binding capacity corresponding to 70 mg IF per 1 kg wet weight. The dried plants still retained 60% of the IF activity. The purified IF preparation consisted of a 50-kDa glycosylated protein with the N-terminal sequence of mature IF. Approximately one-third of the protein was cleaved at the internal site ...PSNPdown arrowGPGP. The key properties of the preparation obtained were identical to those of native IF: the binding curves of vitamin B-12 to recombinant IF and gastric IF were the same, as were those for a B-12 analogue cobinamide, which binds to IF with low affinity. The absorbance spectra of the vitamin bound to recombinant IF and gastric IF were alike, as was the interaction of recombinant and native IF with the specific receptor cubilin. The data presented show that recombinant plants have a great potential as a large-scale source of human IF for analytical and therapeutic purposes.