Abstract
Breast cancer with more than 1.7 million diagnoses per year has been known as one of the most prevalent cancers among women worldwide. Despite the availability of advanced treatment options, cancer progression and metastasis is observed in 20% of patients. Human epidermal growth factor receptor-2 (HER-2) is considered as an important prognostic and diagnostic tumor marker for breast cancer. While HER-2 is expressed on the surface of normal cells, its overexpression occurs in 20–25% on breast cancer tumor cells. This type of tumor which is referred to as HER-2+ is the most aggressive type of breast cancer and shows more resistance to radiotherapy. Single-chain fragment antibodies (ScFvs) offer several advantages in comparison to conventional whole antibodies due to their small size. Particularly, ScFv fragments show improved diffusion and solid tumor penetration. In this study, a human ScFv antibody library was used to isolate anti-HER-2 ScFv antibodies through cell panning and mix antigen-cell panning strategies. Analysis of the binding activity and specificity of isolated ScFv antibodies against HER-2-expressing cell lines and recombinant HER-2 antigen indicated the higher efficiency of the cell panning strategy in isolation of ScFv antibody fragments.





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Ahmad ZA, Yeap SK, Ali AM, Ho WY, Alitheen NBM, and Hamid M (2012) scFv antibody: principles and clinical application. Clinical and Dev Immunol 2012:1–15. https://doi.org/10.1155/2012/980250
Ali SM, Carney WP, Esteva FJ, Fornier M, Harris L, Köstler W, Lotz JP, Luftner D, Pichon MF, Lipton A (2008) Serum HER-2/neu and relative resistance to trastuzumab-based therapy in patients with metastatic breast cancer. Cancer 113(6):1294–1301. https://doi.org/10.1002/cncr.23689
Alirezapour B, Rajabibazl M, Rasaee MJ, Omidfar K (2013) Production and characterization of recombinant scFv against digoxin by phage display technology. Monoclonal Antibodies in Immunodiagnosis and Immunother 32(3):172–179. https://doi.org/10.1089/mab.2012.0093
Ardekani LS, Gargari SLM, Rasooli I, Bazl MR, Mohammadi M, Ebrahimizadeh W, Bakherad H, Zare H (2013) A novel nanobody against urease activity of Helicobacter pylori. Int J Infect Dis 17(9):e723–e728. https://doi.org/10.1016/j.ijid.2013.02.015
Beatty JD, Beatty BG, Vlahos WG (1987) Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Methods 100(1-2):173–179. https://doi.org/10.1016/0022-1759(87)90187-6
Capelan M, Pugliano L, De Azambuja E, Bozovic I, Saini KS, Sotiriou C, Loi S, Piccart-Gebhart MJ (2013) Pertuzumab: new hope for patients with HER2-positive breast cancer. Ann Oncol 24(2):273–282. https://doi.org/10.1093/annonc/mds328
Coussens L, Yang-Feng TL, Liao YC, Chen E, Gray A, McGrath J, Seeburg PH, Libermann TA, Schlessinger J, Francke U (1985) Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science 230(4730):1132–1139. https://doi.org/10.1126/science.2999974
Foroumadi S, Rajabibazl M, Hosseini SH, Rajabi S, Shahidi S, Daraei A, Milani Toofani A (2016) Expression and characterization of recombinant human epidermal growth factor receptor antigene (HER-2) as an indicator of breast cancer in yeast fermented systems. Koomesh 18:110–116
Hosseini H, Rajabibazl M, Ebrahimizadeh W, Dehbidi GR (2015) Inhibiting angiogenesis with human single-chain variable fragment antibody targeting VEGF. Microvasc Res 97:13–18. https://doi.org/10.1016/j.mvr.2014.09.002
Jijakli K, Khraiwesh B, Fu W, Luo L, Alzahmi A, Koussa J, Chaiboonchoe A, Kirmizialtin S, Yen L, Salehi-Ashtiani K (2016) The in vitro selection world. Methods 106:3–13. https://doi.org/10.1016/j.ymeth.2016.06.003
Lennon S, Barton C, Banken L, Gianni L, Marty M, Baselga J, Leyland-Jones B (2009) Utility of serum HER2 extracellular domain assessment in clinical decision making: pooled analysis of four trials of trastuzumab in metastatic breast cancer. J Clin Oncol 27(10):1685–1693. https://doi.org/10.1200/JCO.2008.16.8351
Ma H, O'Kennedy R (2017) Recombinant antibody fragment production. Methods 116:23–33. https://doi.org/10.1016/j.ymeth.2016.11.008
Mohammadzadeh S, Rajabibazl M, Fourozandeh M, Rasaee MJ, Rahbarizadeh F, Mohammadi M (2014) Production of recombinant scFv against p24 of human immunodeficiency virus type 1 by phage display technology. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 33(1):28–33. https://doi.org/10.1089/mab.2013.0059
Moore KM, Thomas GJ, Duffy SW, Warwick J, Gabe R, Chou P, Ellis IO, Green AR, Haider S, Brouilette K, Saha A, Vallath S, Bowen R, Chelala C, Eccles D, Tapper WJ, Thompson AM, Quinlan P, Jordan L, Gillett C, Brentnall A, Violette S, Weinreb PH, Kendrew J, Barry ST, Hart IR, Jones JL, Marshall JF (2014) Therapeutic targeting of integrin alphavbeta6 in breast cancer. J Natl Cancer Inst 106(8) pii:dju169. https://doi.org/10.1093/jnci/dju169
Naume B, Zhao X, Synnestvedt M, Borgen E, Russnes HG, Lingjærde OC, Strømberg M, Wiedswang G, Kvalheim G, Kåresen R (2007) Presence of bone marrow micrometastasis is associated with different recurrence risk within molecular subtypes of breast cancer. Mol Oncol 1(2):160–171. https://doi.org/10.1016/j.molonc.2007.03.004
Olayioye MA (2001) Update on HER-2 as a target for cancer therapy: intracellular signaling pathways of ErbB2/HER-2 and family members. Breast Cancer Res 3(6):385–389. https://doi.org/10.1186/bcr327
Roy V, Perez EA (2009) Beyond trastuzumab: small molecule tyrosine kinase inhibitors in HER-2–positive breast cancer. Oncologist 14(11):1061–1069. https://doi.org/10.1634/theoncologist.2009-0142
Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, Levin WJ, Stuart SG, Udove J, Ullrich A (1989) Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244(4905):707–712. https://doi.org/10.1126/science.2470152
Telli ML, Hunt SA, Carlson RW, Guardino AE (2007) Trastuzumab-related cardiotoxicity: calling into question the concept of reversibility. J Clin Oncol 25(23):3525–3533. https://doi.org/10.1200/JCO.2007.11.0106
Tuefferd M, Couturier J, Penault-Llorca F, Vincent-Salomon A, Broët P, Guastalla J-P, Allouache D, Combe M, Weber B, Pujade-Lauraine E (2007) HER2 status in ovarian carcinomas: a multicenter GINECO study of 320 patients. PLoS One 2(11):e1138. https://doi.org/10.1371/journal.pone.0001138
Wilking U, Karlsson E, Skoog L, Hatschek T, Lidbrink E, Elmberger G, Johansson H, Lindström L, Bergh J (2011) HER2 status in a population-derived breast cancer cohort: discordances during tumor progression. Breast Cancer Res Treat 125(2):553–561. https://doi.org/10.1007/s10549-010-1029-2
Zhang M, Qiu Z, Li Y, Yang Y, Zhang Q, Xiang Q, Su Z, Huang Y (2013) Construction and characterization of a recombinant human beta defensin 2 fusion protein targeting the epidermal growth factor receptor: in vitro study. Appl Microbiol Biotechnol 97(9):3913–3923. https://doi.org/10.1007/s00253-012-4257-z
Acknowledgments
The authors would like to thank the Faculty of Medicine, Shahid Beheshti University of Medical Sciences, for supporting this study. This paper has been extracted from the thesis written by Mrs. Saeideh Foroumadi in School of Medicine, Shahid Beheshti University of Medical Sciences (Registration No: 453).
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Foroumadi, S., Rajabibazl, M., Rahimpour, A. et al. Production of human single-chain fragment antibody (ScFv) against human epidermal growth factor receptor-2 (HER-2) by phage display technology. In Vitro Cell.Dev.Biol.-Animal 54, 85–91 (2018). https://doi.org/10.1007/s11626-017-0221-7
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DOI: https://doi.org/10.1007/s11626-017-0221-7


