Back to Journals » International Journal of Nanomedicine » Volume 6

Design, modeling, expression, and chemoselective PEGylation of a new nanosize cysteine analog of erythropoietin

Authors Ahangari Cohan R, Madadkar-Sobhani A, Khanahmad H, Roohvand F, Aghasadeghi MR, Hedayati MH, Barghi Z, Shafiee Ardestani M, Nouri Inanlou D, Norouzian D

Published 15 June 2011 Volume 2011:6 Pages 1217—1227

DOI https://doi.org/10.2147/IJN.S19081

Review by Single-blind

Peer reviewer comments 3

Reza Ahangari Cohan1, Armin Madadkar-Sobhani2,3, Hossein Khanahmad1, Farzin Roohvand4, Mohammad Reza Aghasadeghi4, Mohammad Hossein Hedayati5, Zahra Barghi5, Mehdi Shafiee Ardestani4, Davoud Nouri Inanlou1, Dariush Norouzian1
1Research and Development Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran; 2Department of Bioinformatics, Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran; 3Department of Life Sciences, Barcelona Supercomputing Center, Barcelona, Spain; 4Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran, Iran; 5Quality Control Department, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran

Background: Recombinant human erythropoietin (rhEPO) is considered to be one of the most pivotal pharmaceutical drugs in the market because of its clinical application in the treatment of anemia-associated disorders worldwide. However, like other therapeutic proteins, it does not have suitable pharmacokinetic properties for it to be administrated at least two to three times per week. Chemoselective cysteine PEGylation, employing molecular dynamics and graphics in in silico studies, can be considered to overcome such a problem.
Methods: A special kind of EPO analog was elicited based on a literature review, homology modeling, molecular dynamic simulation, and factors affecting the PEGylation reaction. Then, cDNA of the selected analog was generated by site-directed mutagenesis and subsequently cloned into the expression vector. The construct was transfected to Chinese hamster ovary/dhfr- cells, and highly expressed clones were selected via methotrexate amplification. Ion-immobilized affinity and size exclusion (SE) chromatography techniques were used to purify the expressed analog. Thereafter, chemoselective PEGylation was performed and a nanosize PEGylated EPO was obtained through dialysis. The in vitro biologic assay and in vivo pharmacokinetic parameters were studied. Finally, E31C analog Fourier transform infrared, analytical SE-high-performance liquid chromatography, zeta potential, and size before and after PEGylation were characterized.
Results: The findings indicate that a novel nanosize EPO31-PEG has a five-fold longer terminal half-life in rats with similar biologic activity compared with unmodified rhEPO in proliferation cell assay. The results also show that EPO31-PEG size and charge versus unmodified protein was increased in a nanospectrum, and this may be one criterion of EPO biologic potency enhancement.
Discussion: This kind of novel engineered nanosize PEGylated EPO has remarkable advantages over rhEPO.
Keywords: nanoPEGylated EPO, cysteine PEGylation, pharmacokinetic property

Creative Commons License This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution - Non Commercial (unported, v3.0) License. By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms.

Download Article [PDF]  View Full Text [HTML][Machine readable]

 

Readers of this article also read:

Degradation and osteogenic potential of a novel poly(lactic acid)/nano-sized β-tricalcium phosphate scaffold

Cao L, Duan PG, Wang HR, Li XL, Yuan FL, Fan ZY, Li SM, Dong J

International Journal of Nanomedicine 2012, 7:5881-5888

Published Date: 28 November 2012

Encapsulation of cisplatin in long-circulating and pH-sensitive liposomes improves its antitumor effect and reduces acute toxicity

Leite EA, Souza CM, Carvalho-Júnior AD, Coelho LG, Lana AM, Cassali GD, Oliveira MC

International Journal of Nanomedicine 2012, 7:5259-5269

Published Date: 9 October 2012

Evaluation of the genotoxicity of cellulose nanofibers

de Lima R, Feitosa LO, Maruyama CR, Barga MA, Yamawaki PC, Vieira IJ, Teixeira EM, Corrêa AC, Mattoso LH, Fraceto LF

International Journal of Nanomedicine 2012, 7:3555-3565

Published Date: 11 July 2012

Lipid nanoparticles as delivery vehicles for the Parietaria judaica major allergen Par j 2

Bondì ML, Montana G, Craparo EF, Di Gesù R, Giammona G, Bonura A, Colombo P

International Journal of Nanomedicine 2011, 6:2953-2962

Published Date: 21 November 2011

PLLA-PEG-TCH-labeled bioactive molecule nanofibers for tissue engineering

Chen J, Zhou B, Li Q, Ouyang J, Kong J, Zhong W, Xing MM

International Journal of Nanomedicine 2011, 6:2533-2542

Published Date: 21 October 2011

Molecular network topology and reliability for multipurpose diagnosis

Jalil MA, Moongfangklang N, Innate K, Mitatha S, Ali J, Yupapin PP

International Journal of Nanomedicine 2011, 6:2385-2392

Published Date: 19 October 2011

Anticancer efficacy enhancement and attenuation of side effects of doxorubicin with titanium dioxide nanoparticles

Chen Y, Wan Y, Wang Y, Zhang H, Jiao Z

International Journal of Nanomedicine 2011, 6:2321-2326

Published Date: 18 October 2011

Phase I dose escalation safety study of nanoparticulate paclitaxel (CTI 52010) in normal dogs

Axiak SM, Selting KA, Decedue CJ, Henry CJ, Tate D, Howell J, Bilof KJ, Kim DY

International Journal of Nanomedicine 2011, 6:2205-2212

Published Date: 11 October 2011

Practical preparation procedures for docetaxel-loaded nanoparticles using polylactic acid-co-glycolic acid

Keum CG, Noh YW, Baek JS, Lim JH, Hwang CJ, Na YG, Shin SC, Cho CW

International Journal of Nanomedicine 2011, 6:2225-2234

Published Date: 7 October 2011