title: Expression, purification, and characterization of recombinant human transferrin from rice (Oryza sativa L.).
abstract: Transferrin is an essential ingredient used in cell culture media due to its crucial role in regulating cellular iron uptake, transport, and utilization. It is also a promising drug carrier used to increase a drug's therapeutic index via the unique transferrin receptor-mediated endocytosis pathway. Due to the high risk of contamination with blood-borne pathogens from the use of human or animal plasma-derived transferrin, recombinant transferrin is preferred for use as a replacement for native transferrin. We expressed recombinant human transferrin in rice (Oryza sativa L.) at a high level of 1% seed dry weight (10 g/kg). The recombinant human transferrin was able to be extracted with saline buffers and then purified by a one step anion exchange chromatographic process to greater than 95% purity. The rice-derived recombinant human transferrin was shown to be not only structurally similar to the native human transferrin, but also functionally the same as native transferrin in terms of reversible iron binding and promoting cell growth and productivity. These results indicate that rice-derived recombinant human transferrin should be a safe and low cost alternative to human or animal plasma-derived transferrin for use in cell culture-based biopharmaceutical production of protein therapeutics and vaccines.
authors.affiliation: Ventria Bioscience, 2860 W Covell Blvd., Suite 1, Davis, CA 95616, United States. DZhang@ventria.com
authors.forename: Deshui
authors.initials: D
authors.lastname: Zhang
authors.forename: Somen
authors.initials: S
authors.lastname: Nandi
authors.forename: Paula
authors.initials: P
authors.lastname: Bryan
authors.forename: Steve
authors.initials: S
authors.lastname: Pettit
authors.forename: Diane
authors.initials: D
authors.lastname: Nguyen
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Santos
authors.forename: Ning
authors.initials: N
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000917:Antibody Formation; D049109:Cell Proliferation; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D007501:Iron / Q000378:metabolism; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014168:Transferrin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D014168:Transferrin; D007501:Iron
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