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Heydari M, Aliabadi Farahani Z, Bayat E, Damough S, Sabzalinezhad M, Talebkhan Garoosi Y. Design, Expression, Purification, and Evaluation of Anti-HER2 scFv. mjms. 2020; 23 (1) :49-56
URL: http://mjms.modares.ac.ir/article-30-42996-en.html
1- Biochemistry Department, Advanced Sciences and Technology Faculty, Medical Sciences Islamic Azad University, Tehran, Iran
2- Biology Department, Science and Research Branch, Islamic Azad University, Tehran, Iran
3- Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
4- Biotechnology Department, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University
5- Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran , talebkhan@pasteur.ac.ir
Abstract:   (943 Views)
Aims: The aim of the present study was to design and express an anti-HER2 single chain variable antibody fragment in E. coli BL21 (DE3) and evaluate its efficiency in recognition of HER2 protein.
Materials & Methods: An approximately 746bp encoding gene fragment was cloned into pET28a and the recombinant protein was expressed in E. coli BL21 (DE3) strain. Following protein purification by affinity chromatography, western blotting and ELISA were used to evaluate the efficiency of anti-HER2 scFv against HER2 protein.
Findings: E. coli can express the anti-HER2 scFv molecule possessing appropriate function and can detect this protein on the surface of breast cancer cells.
Conclusion: This antibody fragment can be used in laboratory diagnostic methods for HER2 diagnostic approaches. Potential capability of this protein in immunohistochemical and imaging approaches against HER2 should be considered.
Full-Text [PDF 507 kb]   (278 Downloads)    
Article Type: Original Research | Subject: Biotechnology
Received: 2020/05/19 | Accepted: 2020/06/11

1. Cho HS, Mason K, Ramyar KX, Stanley AM, Gabelli SB, Denney DW, et al. Structure of the extracellular region of HER2 alone and in complex with the Herceptin Fab. Nature. 2003;421(6924):756-60. [Link] [DOI:10.1038/nature01392]
2. Weisser NE, Hall JC. Applications of single-chain variable fragment antibodies in therapeutics and diagnostics. Biotechnol Adv. 2009;27(4):502-20. [Link] [DOI:10.1016/j.biotechadv.2009.04.004]
3. Galeffi P, Lombardi A, Pietraforte I, Novelli F, Di Donato M, Sperandei M, et al. Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems. J Transl Med. 2006;4(1):39. [Link] [DOI:10.1186/1479-5876-4-39]
4. Sommaruga S, Lombardi A, Salvadè A, Mazzucchelli S, Corsi F, Galeffi P, et al. Highly efficient production of anti-HER2 scFv antibody variant for targeting breast cancer cells. Appl Microbiol Biotechnol. 2011;91(3):613-21. [Link] [DOI:10.1007/s00253-011-3306-3]
5. Rostinawati T, Kusuma SA, Yusuf M, Gaffar S, Subroto T. Construction and Expression of Synthetic-gene encoding anti-HER2 scFv Fused with pelB in Escherichia coli BL21 (DE3). J Pharm Sci Res. 2017;9(11):2106-13. [Link]
6. Farshdari F, Ahmadzadeh M, Jahandar H, Mohit E. Enhanced solubility of anti-HER2 scFv using bacterial Pelb leader sequence. Iran J Pharm Sci. 2019;15(1):1-6. [Link]
7. Farshdari F, Ahmadzadeh M, Nematollahi L, Mohit E. The improvement of anti-HER2 scFv soluble expression in Escherichia coli. Braz J Pharm Sci. 2020;56. [Link] [DOI:10.1590/s2175-97902019000317861]
8. Cheng WW, Allen TM. The use of single chain Fv as targeting agents for immunoliposomes: An update on immunoliposomal drugs for cancer treatment. Expert Opin Drug Deliv. 2010;7(4):461-78. [Link] [DOI:10.1517/17425240903579963]
9. Marks JD, Poul MA, inventors. Internalizing ErbB2 antibodies. WIPO (PCT) patent WO1999055367A1. 1999. [Link]
10. Pedersen MW, Jensen A, Meijer P, inventors. Anti-HER2 antibodies and compositions. United States patent US8609095B2. 2011. [Link]
11. Thongsuksai P, Chongsuvivatwong V, Sriplung H. Delay in breast cancer care: A study in Thai women. Med Care. 2000;38(1):108-14. [Link] [DOI:10.1097/00005650-200001000-00012]
12. Arteaga CL, Sliwkowski MX, Osborne CK, Perez EA, Puglisi F, Gianni L. Treatment of HER2-positive breast cancer: Current status and future perspectives. Nat Rev Clin Oncol. 2012;9(1):16-32. [Link] [DOI:10.1038/nrclinonc.2011.177]
13. Ahmadi A, Mohagheghi MA, Fazeli MS, Nahavandian B, Bashardoost N, Jarahi AM, et al. HESA-A: New treatment for breast cancer and choroidal metastasis. Med Sci Monit. 2005;11(6):CR300-3. [Link]
14. Murphy CG, Modi Sh. HER2 breast cancer therapies: A review. Biologics. 2009;3:289-301. [Link] [DOI:10.2147/BTT.S3479]
15. Tai W, Mahato R, Cheng K. The role of HER2 in cancer therapy and targeted drug delivery. J Controll Release. 2010;146(3):264-75. [Link] [DOI:10.1016/j.jconrel.2010.04.009]
16. Akbari V, Sadeghi HM, Jafrian-Dehkordi A, Abedi D, Chou CP. Functional expression of a single-chain antibody fragment against human epidermal growth factor receptor 2 (HER2) in Escherichia coli. J Ind Microbiol Biotechnol. 2014;41(6):947-56. [Link] [DOI:10.1007/s10295-014-1437-0]
17. Jaliani HZ, Farajnia S, Safdari Y, Mohammadi SA, Barzegar A, Talebi S. Optimized condition for enhanced soluble-expression of recombinant mutant anabaena variabilis phenylalanine ammonia lyase. Adv Pharm Bull. 2014;4(3):261-6. [Link]
18. Naderi S, Alikhani MY, Karimi J, Shabab N, Mohamadi N, Jaliani HZ, et al. Cytoplasmic expression, optimization and catalytic activity evaluation of recombinant mature lysostaphin as an anti-staphylococcal therapeutic in escherichia coli. Acta Med Int. 2015;2(2):72-7. [Link] [DOI:10.5530/ami.2015.3.5]
19. Napathorn SC, Kuroki M, Kuroki M. High expression of fusion proteins consisting of a single-chain variable fragment antibody against a tumor-associated antigen and interleukin-2 in Escherichia coli. Anticancer Res. 2014;34(8):3937-46. [Link]
20. Kim S, Cheung LH, Zhang W, Rosenblum MG. Improved expression of a soluble single chain antibody fusion protein containing tumor necrosis factor in Escherichia coli. Appl Microbiol Biotechnol. 2007;77(1):99-106. [Link] [DOI:10.1007/s00253-007-1133-3]
21. Huang R, Wang Q, Zhang X, Zhu J, Sun B. Trastuzumab-cisplatin conjugates for targeted delivery of cisplatin to HER2-overexpressing cancer cells. Biomed Pharmacother. 2015;72:17-23. [Link] [DOI:10.1016/j.biopha.2015.03.004]

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