N-Linked Glycan Structures of Human Lactoferrin Produced by Transgenic Rice

A Calorimetric Study of Thermal Denaturation of Recombinant Human Lactoferrin from Rice

“…There are also differences in the type of glycans due to the different mechanisms of glycosylation that the vegetable cells have as compared to those of the mammary gland epithelial cells. Thus, lactoferrin from human milk has the typical glycans of mammals, such as R2-6-linked neuraminic acid, β1-4-linked galactose, and R1-6-linked fucose (32), and lactoferrin from rice has the typical glycans of vegetables such as R1-3-linked fucose and β1-2-linked xilose (14). Differences in glycosylation have also been found in other rhLFs such as those produced in A. awamori (8), in the milk of transgenic cows (9), or in the milk of transgenic mice (33).…”

“…It is important to determine the thermal stability of rhLF to design treatments that ensure that the structure and biological activity of the protein are maintained. Although it has been reported that rhLF from rice possesses similar biological activities and a similar structure to human milk lactoferrin (12,13), there are differences in the glycosylation pattern, which could be important in the behavior of the protein (14). Differences have also been reported in the glycans of the different transferrins (15) and also in the lactoferrin isolated from leucocytes as compared with that from milk (16).…”

Plant‐based biopharming of recombinant human lactoferrin

“…In many countries, rice grains are the first solid baby food due to rice hypoallergenicity and commercial availability (Nandi et al, 2002). Native Lf has a2-6-linked neuraminic acid, b1-4-linked galactose and a1-6-linked fucose glycanstypical for mammals (Spik et al, 1982), while rhLf from "koshihikari" rice cultivar has a1-3-linked fucose and b1-2-linked xylosetypical plant glycans (Fujiyama et al, 2004). Additionally, the promoter shows specifically unregulated activity during seed maturation (Huang et al, 2010) and codon optimization (Nandi et al, 2002;Suzuki et al, 2003;Rachmawati et al, 2005;Huang et al, 2010;Lee et al, 2010;Lin et al, 2010), which are extra benefits of this rice-based expression system.…”

“…Codon optimization by the replacement of either adenine or thymine at the third position to cytosine or guanine resulted in the increase of the recombinant hLf production level from 0.5 to 5.0 g/kg in the dehusked rice grains. Native Lf has a2-6-linked neuraminic acid, b1-4-linked galactose and a1-6-linked fucose glycanstypical for mammals (Spik et al, 1982), while rhLf from "koshihikari" rice cultivar has a1-3-linked fucose and b1-2-linked xylosetypical plant glycans (Fujiyama et al, 2004).…”

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2003–2004

“…(Hoshi, et al, 2004) IgG (Rituximab, Trastuzumab) Commercial PNGase F TOF (3-AA), CE Method for monitoring glycosylation in pharmaceuticals based on capillary electrophoresis with structural confirmation by MALDI-TOF MS (Kamoda, et al, 2004) IgG (Sriraman, et al, 2004) IgG 1 (Human) Rat hybridoma or CHO Hydrazine MALDI, HPLC, Glycans (2-AP), Absence of fucose but not presence of galactose or bisect enhances cellular toxicity (Shinkawa, et al, 2003) IgG 3 M o u s e Hydrazine, PNGase F R-TOF (+ve) (DHB), ESI, MS/MS, Glycans Effect of buffer and pH on production. (Müthing, et al, 2003) IgG 4 (Human) CHO-S cells Trypsin TOF (CHCA), Glycopeptides Glycosylation (high-mannose, truncated complex) not significantly affected by anti-mitotic agent (Tait, et al, 2004) KDEL-tagged cPIPP antibody Structural characterization (Fujiyama, et al, 2004) Lecithin:cholesterol acyltransferase (Lane, et al, 2004) Luteinizing hormone (Human) CHO cells Hydrazine R -TOF (DHB, CMBT), ESI, Ion trap, Glycans (2-AB) Structural determination of complex glycans and method development (Gervais, et al, 2003) MFE-CP…”