Optimization of HER2-based and cell-based ELISA for detection of trastuzumab biosimilar

authors:

avatar Maryam Ahmadzadeh , avatar Farzaneh Farshdari , avatar Leila Nematollahi , avatar Shayan Maleknia , avatar Elham Mohit , *


how to cite: Ahmadzadeh M, Farshdari F, Nematollahi L, Maleknia S, Mohit E. Optimization of HER2-based and cell-based ELISA for detection of trastuzumab biosimilar. koomesh. 2020;22(2):e153187. 

Abstract

Introduction: ELISA is a sensitive, specific, reproducible and fast method to quantify the biological activity of antibodies. Trastuzumab is a humanized monoclonal antibody against HER2 receptors which prevents the initiation of downstream signaling pathway. Trastuzumab can be used as a positive control in the ELISA experiments for anti-HER2 antibodies. Additionaly, insufficient washing and blocking can be the cause of background in the ELISA experiment that is necessary to be optimized. As we do not want to waste antigen, it is also important to determine the last dilution of antigen that can give good titration curve for trastuzumab. The aim of this study was to optimize ELISA method for trastuzumab biosimilar (AryoTrust™, Aryogen pharmed). Materials and Methods: In this study, different variables including the number of washing cycle, the blocking agent and antigen concentration in HER2-based ELISA and the type of HER2 negative cell line and the blocking agent in cell-based ELISA were studied. Results: It was demonstrated that 5 times washing between different steps of HER2-based ELISA causes significant lower non-specific background as compared to 3 times washing. Moreover, the nonspecific binding was significantly lower in the presence of % 5 skim milk as blocking agent as compared to BSA %1 or BSA %3. In addition, the lowest HER2 concentration which gives good titration curve for trastuzumab was 0.1 μg/ml. In cell-based ELISA experiment, it was demonstrated that the use of MDA-MB-231 as negative HER2 cell line caused significant lower background than MCF-7. Furthermore, BSA 3% was chosen as proper blocking agent. Conclusion: The results of this study can be used for development of HER2 and cell-based ELISA for anti HER2 antibodies and its fragments.

References

  • 1.

    Thongsuksai P, Chongsuvivatwong V, Sriplung H. Delay in breast cancer care: a study in Thai women. Med Care 2000; 38: 108-114.

  • 2.

    Taslimi Y, Zahedifard F, Habibzadeh S, Taheri T, Abbaspour H, Sadeghipour A, et al. Antitumor effect of IP-10 by using two different approaches: live delivery system and gene therapy. J Breast Cancer 2016; 19: 34-44.

  • 3.

    Semnani V, Farhidzadeh E, Mirmohammadkhani M, Ghahremanfard F. Investigation of blood levels of vitamin D in women with breast cancer and its correlation with prognostic markers. Koomesh 2017; 735-741. (Persian).

  • 4.

    Parise CA, Bauer KR, Brown MM, Caggiano V. Breast cancer subtypes as defined by the estrogen receptor (ER), progesterone receptor (PR), and the human epidermal growth factor receptor 2 (HER2) among women with invasive breast cancer in California, 1999-2004. Breast J 2009; 15: 593-602.

  • 5.

    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: 16-32.

  • 6.

    Ahmadi A, Mohagheghi MA, Fazeli MS, Nahavandian B, Bashardoost N, Jarahi AM, Gharipoor M. HESA-A: new treatment for breast cancer and choroidal metastasis. Med Sci Monit 2005; 11: CR300-CR303.

  • 7.

    Murphy CG, Modi S. HER2 breast cancer therapies: a review. Biologics 2009; 3: 289-301.

  • 8.

    Kauraniemi P, Kallioniemi A. Activation of multiple cancer-associated genes at the ERBB2 amplicon in breast cancer. Endocr Relat Cancer 2006; 13: 39-49.

  • 9.

    Foroumadi S, Rajabi bazl M, Hosseini H, Rajabi S, Shahidi S, Daraei A, Toofani Milani A. Expression and characterization of recombinant human epidermal growth factor receptor antigene (HER-2) as an indicator of breast cancer in yeast fermented systems. Koomesh 2016; 18: 110-116. (Persian).

  • 10.

    Tai W, Mahato R, Cheng K. The role of HER2 in cancer therapy and targeted drug delivery. J Control Release 2010; 146: 264-275.

  • 11.

    Mohit E, Hashemi A, Allahyari M. Breast cancer immunotherapy: monoclonal antibodies and peptide-based vaccines. Expert Rev Clin Immunol 2014; 10: 927-961.

  • 12.

    Surez I, Salmern-Garca A, Cabeza J, Capitn-Vallvey LF, Navas N. Development and use of specific ELISA methods for quantifying the biological activity of bevacizumab, cetuximab and trastuzumab in stability studies. J Chromatogr B Analyt Technol Biomed Life Sci 2016; 1032: 155-164.

  • 13.

    Cheng W, Allen T. 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: 461-478.

  • 14.

    Damen CW, de Groot ER, Heij M, Boss DS, Schellens JH, Rosing H, et al. Development and validation of an enzyme-linked immunosorbent assay for the quantification of trastuzumab in human serum and plasma. Anal Biochem 2009; 391: 114-120.

  • 15.

    Steinitz M. Quantitation of the blocking effect of tween 20 and bovine serum albumin in ELISA microwells. Anal Biochem 2000; 282: 232-238.

  • 16.

    Sentandreu MA, Aubry L, Toldr F, Ouali A. Blocking agents for ELISA quantification of compounds coming from bovine muscle crude extracts. Eur Food Res Technol 2007; 224: 623-628.

  • 17.

    Khan MS, Pande T, van de Ven TG. Qualitative and quantitative detection of T7 bacteriophages using paper based sandwich ELISA. Colloids Surf B Biointerfaces 2015; 132: 264-270.

  • 18.

    Crowther R. The ELISA Guidebook. ed. Humana Press; 2009; 88-97.

  • 19.

    Mosayebi G, Abtahi H, Ghazavi A, Zareinfar N, Khaki M, Payani MA. Design of enzyme-linked immunosorbent assay method for detection of anti-streptolysin-O antibodies on base of recombinant streptolysin-O. Koomesh 2012; 13: 362-368. (Persian).

  • 20.

    Pterfi Z, Kocsis B. Comparison of blocking agents for an ELISA for LPS. J Immunoassay 2000; 21: 341-354.

  • 21.

    Coory M, Thornton K. Randomised clinical endpoint studies for trastuzumab biosimilars: a systematic review. Breast Cancer Res Treat 2019; 1: 17-25.

  • 22.

    Holbech H, Andersen L, Petersen GI, Korsgaard B, Pedersen KL, Bjerregaard P. Development of an ELISA for vitellogenin in whole body homogenate of zebrafish (Danio rerio). Comp Biochem Physiol C Toxicol Pharmacol 2001; 130: 119-131.

  • 23.

    Dixit CK, Vashist SK, MacCraith BD, O'kennedy R. Multisubstrate-compatible ELISA procedures for rapid and high-sensitivity immunoassays. Nat Protoc 2011; 6: 439-445.

  • 24.

    Kaur R, Dikshit KL, Raje M. Optimization of immunogold labeling TEM: an ELISA-based method for evaluation of blocking agents for quantitative detection of antigen. J Histochem Cytochem 2002; 50: 863-873.

  • 25.

    Khodashenas S, Khalili S, Moghadam MF. A cell ELISA based method for exosome detection in diagnostic and therapeutic applications. Biotechnol Lett 2019; 41: 523-531.

  • 26.

    Meysami P, Rezaei F, Marashi SM, Amiri MM, Bakker E, Mokhtari-Azad T. Antitumor effects of a recombinant baculovirus displaying anti-HER2 scFv expressing Apoptin in HER2 positive SK-BR-3 breast cancer cells. Future Virol 2019; 14: 139-152.

  • 27.

    Maple L, Lathrop R, Bozich S, Harman W, Tacey R, Kelley M, Danilkovitch-Miagkova A. Development and validation of ELISA for Herceptin detection in human serum. J Immunol Methods 2004; 295: 169-182.

  • 28.

    Tan WB, Jiang S, Zhang Y. Quantum-dot based nanoparticles for targeted silencing of HER2/neu gene via RNA interference. Biomaterials 2007; 28: 1565-1571.

  • 29.

    Alric C, Herv-Aubert K, Aubrey N, Melouk S, Lajoie L, Mme W, et al. Targeting HER2-breast tumors with scFv-decorated bimodal nanoprobes. J Nanobiotechnology 2018; 16: 18.

  • 30.

    Lattrich C, Juhasz-Boess I, Ortmann O, Treeck O. Detection of an elevated HER2 expression in MCF-7 breast cancer cells overexpressing estrogen receptor 1. Oncol Rep 2008; 19: 811-817.

  • 31.

    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.##https://doi.org/10.1016/j.biopha.2015.03.004.

  • 32.

    Sarial S, Sonboli R, Maleknia S, Ashori F, Rezaeiravesh S, Nouri SZ, et al. Development, optimization and validation of a sandwich ELISA for detection of recombinant human factor VII. South Asian J Exper Biol 2015; 5: 26-31.##.