Abstract
In this paper, we show that recombinant human lactoferrin (rhLF) has been stably expressed at 0.5% brown rice flour weight for nine generations. Process development indicates that rhLF can be efficiently extracted from rice flour in 20 mM phosphate buffer (pH 7.0) containing up to 0.5 M NaCl and at a ratio of 1 kg flour to 10 L buffer. After solid/liquid separation, the extract can then be loaded directly onto an ion-exchange column and rhLF can be eluted using 0.8 M NaCl. The resulting rhLF is about 95 pure. A range of biochemical and biophysical analyses were carried out and results indicated that the purified rhLF was identical to its native human counterpart other than its glycosylation. Economic analysis shows that at 600 kg/year scale, the cash cost to produce 1 g of rhLF of pharmaceutical grade is US$ 5.90. Analysis also indicates that the expression level has profound impact on costs related to planting, milling, extraction and purification, thus high level expression of recombinant protein in plants is one of the key parameters for the success of plant made pharmaceuticals.
Similar content being viewed by others
References
Anzai H, Takaiwa F and Katsumata K (1999) Production of human lactoferrin in transgenic plants. In: Shimazaki K, Tsuda H, Tomita M, Kuwata T and Perraudin J-P (eds.), International Congress on Lactoferrin: Structure, Function and Application. (pp 265–271). Elsevier Science B.V., Burgerhartstraat, Sapporo, Japan, 18–22 May. 1999.
MK Bhan NK Arora V Khoshoo P Raj S Bhatnager S Sazawal K Sharma (1988) ArticleTitleComparison of a lactose-free cereal-based formula and cow’s milk in infants and children with acute gastroenteritis J Pediatr Gastroenterol Nutr 7 208–213 Occurrence Handle3351705
DK Chong WH Langridge (2000) ArticleTitleExpression of full-length bioactive antimicrobial human lactoferrin in potato plants Transgenic Research 9 71–78 Occurrence Handle10853271
C Christy G Adams R Kuriyel G Bolton A Seilly (2002) ArticleTitleHigh-performance tangential flow filtration: A highly selective membrane seperation process Desalination 144 133–136
H Daniell SJ Streatfield K Wycoff (2001) ArticleTitleMedical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants Trends Plant Sci 6 219–226 Occurrence Handle11335175
RL Evangelista AR Kusnadi JA Howard ZL Nikolov (1998) ArticleTitleProcess and economic evaluation of the extraction and purification of recombinant beta-glucuronidase from transgenic corn Biotechnol Prog 14 607–614 Occurrence Handle9694683
A Gastañaduy A Cordano GG Graham (1990) ArticleTitleAcceptability, tolerance, and nutritional value of a rice-based infant formula J Pediatric Gastroenterol Nutr 11 240–246
RM Helm AW Burks (2000) ArticleTitleMechanisms of food allergy Curr Opin Immunol 12 647–653 Occurrence Handle11102767
EE Hood SL Woodard (2002) Industrial proteins produced from transgenic plants EE Hood JA Howard (Eds) Plants as Factories for Protein Production. Kluwer Academic Publishers The Netherlands 119–135
N Huang (2004) ArticleTitleHigh-level protein expression system uses self-pollinating crops as host BioProcess International 2 IssueID(1 54–59
BD Humphrey N Huang KC Klasing (2002) ArticleTitleRice expressing lactoferrin and lysozyme has antibiotic-like properties when fed to chicks J Nutr 132 1214–1218 Occurrence Handle12042436
B Lönnerdal (1996) ArticleTitleRecombinant human milk proteins - an opportunity and a challenge Am J Clin Nut 63 622S–626S
S Matsumoto K Ikura M Ueda R Sasaki (1995) ArticleTitleCharacterization of a human glycoprotein (erythropoietin) produced in cultured tobacco cells Plant Mol Biol 27 1163–1172 Occurrence Handle7766897
J Messeguer C Fogher E Guiderdoni V Marfa MM Catala G Baldi E Mele (2001) ArticleTitleField assessment of gene flow from transgenic to cultivated rice (Oryza sativa L.) using a herbicide resistance gene as tracer marker Theor Appl Genet 103 1151–1159
D Mison J Curling (2000) ArticleTitleThe industrial production cost of recombinant therapeutic protein expressed in transgenic corn Biopharm Int 13 48–54
A Mitra Z Zhang (1994) ArticleTitleExpression of a human lactoferrin cDNA in tobacco cells produces antibacterial protein(s) Plant Physiol 106 977–981 Occurrence Handle7824662
S Nandi A Suzuki J Huang D Yalda P Pham L Wu G Bartley N Huang B Lonnerdal (2002) ArticleTitleExpression of human lactoferrin in transgenic rice grains for the application in infant formula Plant Science 163 713–722
InstitutionalAuthorNameNIH publication (1984) American Academy of Allergy and Immunology committee on Adverse Reaction to Food and National Institute of Allergy and Infectious Diseases National Institute of Health Bethesda 130–131
MS Peters KD Timmerhaus (1990) Plant Design and Economics for Chemical Engineers EditionNumber4 McGraw-Hill New York
D Prtridis E Sapidou J Calandranis (1995) ArticleTitleComputer-aided process analysis and economic evaluation for biosynthetic human insulin production-a study case Biotechnol Bioeng 48 529–541
AS Rathore GV Johnson JJ Buckley DM Boyle ME Gustafson (2003) ArticleTitleProcess characterization of the chromatographic steps in the purification process of a recombinant Escherichia coli-expressed protein Biotechnol Appl Biochem 37 51–61 Occurrence Handle12578552
AS Rathore P Latham H Levine J Curling O Kaltenbrunner (2004) ArticleTitleCosting issues in the production of biopharmaceuticals Biopharm Int 17 46–55
A Sadana AM Beelaram (1994) ArticleTitleEfficiency and economics of bioseparation: some case studies Bioseparation 4 221–235 Occurrence Handle7765088
V Salmon D Legrand MC Slomianny I ElYazidi G Spik V Gruber P Bournat B Olagnier D Mison M Theisen B Merot (1998) ArticleTitleProduction of human lactoferrin in transgenic tobacco plants Protein Exp Purif 13 127–135
J Schwab (2003) A strategic evaluation of transgenic plant and animal biomanufacturing systems Revelogic, Inc. Fort Collins, CO
L Schwartz (2003) ArticleTitleDiafiltration for Desalting or Buffer Exchange BioProcess International 1 IssueID5 43–49
AB Schyver (1989) ArticleTitleIdentification of transferrin- and lactoferrin-binding proteins in Haemophilus influenzae J Med Microbiol 29 121–130 Occurrence Handle2543820
FF Shih (2003) ArticleTitleAn update on the processing of high-protein rice products Nahrung 47 420–424 Occurrence Handle14727771
K-C Shimazaki H Tsuda M Tomita T Kuwata P J-P. eds (2000) Lactoferrin: Structure, Function and Application Elsevier Amsterdam
RM Twyman E Stoger S Schillberg P Christou R Fischer (2003) ArticleTitleMolecular farming in plants: host systems and expression technology Trends Biotechnol 21 570–578 Occurrence Handle14624867
PH Berkel Particlevan MM Welling M Geerts HA Veen Particlevan B Ravensbergen M Salaheddine EK Pauwels F Pieper JH Nuijens PH Nibbering (2002) ArticleTitleLarge scale production of recombinant human lactoferrin in the milk of transgenic cows Nat Biotechnol 20 484–487 Occurrence Handle11981562
PP Ward JY Lo M Duke GS May DR Headon OM Conneely (1992) ArticleTitleProduction of biologically active recombinant human lactoferrin in Aspergillus oryzae Bio Technol 10 784–789
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nandi, S., Yalda, D., Lu, S. et al. Process development and economic evaluation of recombinant human lactoferrin expressed in rice grain. Transgenic Res 14, 237–249 (2005). https://doi.org/10.1007/s11248-004-8120-6
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11248-004-8120-6