Full Text:   <2206>

CLC number: Q943.2

On-line Access: 2013-10-08

Received: 2013-03-26

Revision Accepted: 2013-06-06

Crosschecked: 2013-09-22

Cited: 4

Clicked: 3122

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE B 2013 Vol.14 No.10 P.867-874


Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice

Author(s):  Qing Zhang, Hui Yu, Feng-zhen Zhang, Zhi-cheng Shen

Affiliation(s):  State Key Laboratory of Rice Biology and Institute of Insect Sciences, Zhejiang University, Hangzhou 310058, China

Corresponding email(s):   zcshen@zju.edu.cn

Key Words:  Recombinant human serum albumin (rHSA), Selectively terminable transgenic rice, Purification

Share this article to: More |Next Article >>>

Qing Zhang, Hui Yu, Feng-zhen Zhang, Zhi-cheng Shen. Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice[J]. Journal of Zhejiang University Science B, 2013, 14(10): 867-874.

@article{title="Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice",
author="Qing Zhang, Hui Yu, Feng-zhen Zhang, Zhi-cheng Shen",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice
%A Qing Zhang
%A Hui Yu
%A Feng-zhen Zhang
%A Zhi-cheng Shen
%J Journal of Zhejiang University SCIENCE B
%V 14
%N 10
%P 867-874
%@ 1673-1581
%D 2013
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300090

T1 - Expression and purification of recombinant human serum albumin from selectively terminable transgenic rice
A1 - Qing Zhang
A1 - Hui Yu
A1 - Feng-zhen Zhang
A1 - Zhi-cheng Shen
J0 - Journal of Zhejiang University Science B
VL - 14
IS - 10
SP - 867
EP - 874
%@ 1673-1581
Y1 - 2013
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1300090

Human serum albumin (HSA) is widely utilized for medical purposes and biochemical research. Transgenic rice has proved to be an attractive bioreactor for mass production of recombinant HSA (rHSA). However, transgene spread is a major environmental and food safety concern for transgenic rice expressing proteins of medical value. This study aimed to develop a selectively terminable transgenic rice line expressing HSA in rice seeds, and a simple process for recovery and purification of rHSA for economical manufacture. An HSA expression cassette was inserted into a T-DNA vector encoding an RNA interference (RNAi) cassette suppressing the CYP81A6 gene. This gene detoxifies the herbicide bentazon and is linked to the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) cassette which confers glyphosate tolerance. ANX Sepharose Fast Flow (ANX FF) anion exchange chromatography coupled with Butyl Sepharose High Performance (Butyl HP) hydrophobic interaction chromatography was used to purify rHSA. A transgenic rice line, HSA-84, was obtained with stable expression of rHSA of up to 0.72% of the total dry weight of the dehusked rice seeds. This line also demonstrated high sensitivity to bentazon, and thus could be killed selectively by a spray of bentazon. A two-step chromatography purification scheme was established to purify the rHSA from rice seeds to a purity of 99% with a recovery of 62.4%. Results from mass spectrometry and N-terminus sequencing suggested that the purified rHSA was identical to natural plasma-derived HSA. This study provides an alternative strategy for large-scale production of HSA with a built-in transgene safety control mechanism.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]Belew, M., Yan, M., Zhang, W., Caldwell, K., 2008. Purification of recombinant human serum albumin (rHSA) produced by genetically modified Pichia pastoris. Separ. Sci. Technol., 43(11-12):3134-3153.

[2]Bosse, D., Praus, M., Kiessling, P., Nyman, L., Andresen, C., Waters, J., Schindel, F., 2005. Phase I comparability of recombinant human albumin and human serum albumin. J. Clin. Pharmacol., 45(1):57-67.

[3]Bradford, M.M., 1976. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal. Biochem., 72(1-2):248-254.

[4]Cai, C., Zhou, K., Wu, Y., Wu, L., 2006. Enhanced liver targeting of 5-fluorouracil using galactosylated human serum albumin as a carrier molecule. J. Drug Target., 14(2):55-61.

[5]Chuang, V.T.G., Otagiri, M., 2007. Recombinant human serum albumin. Drugs Today, 43(8):547-561.

[6]Dong, Y., Zhang, F., Wang, Z., Du, L., Hao, A., Jiang, B., Tian, M., Li, Q., Jia, Q., Wang, S., et al., 2012. Extraction and purification of recombinant human serum albumin from Pichia pastoris broths using aqueous two-phase system combined with hydrophobic interaction chromatography. J. Chromatogr. A, 1245:143-149.

[7]Erstad, B.L., 1996. Viral infectivity of albumin and plasma protein fraction. Pharmacotherapy, 16(6):996-1001.

[8]Farran, I., Sánchez-Serrano, J., Medina, J., Prieto, J., Mingo-Castel, A., 2002. Targeted expression of human serum albumin to potato tubers. Transgenic Res., 11(4):337-346.

[9]Fernández-San Millán, A., Mingo-Castel, A., Miller, M., Daniell, H., 2003. A chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnol. J., 1(2):71-79.

[10]Fox, J.L., 2003. Puzzling industry response to ProdiGene fiasco. Nat. Biotechnol., 21(1):3-4.

[11]Hammitt, D., Walker, D., Syrop, C., Miller, T., Bennett, M., 1991. Treatment of severe male-factor infertility with high concentrations of motile sperm by microinsemination in embryo cryopreservation straws. J. In Vitro Fert. Embryo Transf., 8(2):101-110.

[12]Hastings, G.E., Wolf, P.G., 1992. The therapeutic use of albumin. Arch. Fam. Med., 1(2):281-287.

[13]He, Y., Ning, T., Xie, T., Qiu, Q., Zhang, L., Sun, Y., Jiang, D., Fu, K., Yin, F., Zhang, W., et al., 2011. Large-scale production of functional human serum albumin from transgenic rice seeds. PNAS, 108(47):19078-19083.

[14]Hiei, Y., Ohta, S., Komari, T., Kumashiro, T., 1994. Efficient transformation of rice (Oryza sativa L.) mediated by agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J., 6(2):271-282.

[15]Huang, L.F., Liu, Y.K., Lu, C.A., Hsieh, S.L., Yu, S.M., 2005. Production of human serum albumin by sugar starvation induced promoter and rice cell culture. Transgenic Res., 14(5):569-581.

[16]Huang, N., Rodriquez, R.L., Hagie, F.E., Stalker, D.M., 2007. Expression of Human Serum Albumin (HSA) in Monocot Seeds. US 07304208.

[17]Kobayashi, K., 2006. Summary of recombinant human serum albumin development. Biologicals, 34(1):55-59.

[18]Kratz, F., 2008. Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. J. Control. Release, 132(3):171-183.

[19]Kusnadi, A.R., Nikolov, Z.L., Howard, J.A., 1997. Production of recombinant proteins in transgenic plants: practical considerations. Biotechnol. Bioeng., 56(5):473-484.

[20]Langer, K., Balthasar, S., Vogel, V., Dinauer, N., von Briesen, H., Schubert, D., 2003. Optimization of the preparation process for human serum albumin (HSA) nanoparticles. Int. J. Pharm., 257(1-2):169-180.

[21]Ledford, H., 2007. Out of bounds. Nature, 445(7124):132-133.

[22]Lin, C.Y., Fang, J., Xu, X.L., Zhao, T., Cheng, J.A., Tu, J.M., Ye, G.Y., Shen, Z.C., 2008. A built-in strategy for containment of transgenic plants: creation of selectively terminable transgenic rice. PloS One, 3(3):e1818.

[23]Marth, E., Kleinhappl, B., 2001. Albumin is a necessary stabilizer of TBE-vaccine to avoid fever in children after vaccination. Vaccine, 20(3-4):532-537.

[24]Mendez, C.M., Mcclain, C.J., Marsano, L.S., 2005. Albumin therapy in clinical practice. Nutr. Clin. Pract., 20(3):314-320.

[25]Ohtani, W., Nawa, Y., Takeshima, K., Kamuro, H., Kobayashi, K., Ohmura, T., 1998. Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris. Anal. Biochem., 256(1):56-62.

[26]Pan, G., Zhang, X.Y., Liu, K.D., Zhang, J.W., Wu, X.Z., Zhu, J., Tu, J.M., 2006. Map-based cloning of a novel rice cytochrome P450 gene CYP81A6 that confers resistance to two different classes of herbicides. Plant Mol. Biol., 61(6):933-943.

[27]Subramanian, G.M., Fiscella, M., Lamouse-Smith, A., Zeuzem, S., Mchutchison, J.G., 2007. Albinterferon [α]-2b: a genetic fusion protein for the treatment of chronic hepatitis C. Nat. Biotechnol., 25(12):1411-1419.

[28]Sumi, A., Okuyama, K., Kobayashi, K., Ohtani, W., Ohmura, T., Yokoyama, K., 1999. Purification of recombinant human serum albumin efficient purification using streamline. Bioseparation, 8(1/5):195-200.

[29]Tsuchida, E., Sou, K., Nakagawa, A., Sakai, H., Komatsu, T., Kobayashi, K., 2009. Artificial oxygen carriers, hemoglobin vesicles and albumin-hemes, based on bioconjugate chemistry. Bioconjug. Chem., 20(8):1419-1440.

[30]Vermij, P., 2006. Liberty link rice raises specter of tightened regulations. Nat. Biotechnol., 24(11):1301-1302.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - Journal of Zhejiang University-SCIENCE