GENERATION OF HUMANIZED SINGLE CHAIN ANTI-CD20ANTIBODY MARKER IN E. COLI

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AHMADZADEH V.; FARAJNIA S.; HOSSEINPOUR FEIZI M.A.; KHAVARINEJAD R.A.

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Journal: Journal of Advances in Medical and Biomedical Research Year:2014 | Volume:21 | Issue:88 Page(s): 12-21

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Title

GENERATION OF HUMANIZED SINGLE CHAIN ANTI-CD20ANTIBODY MARKER IN E. COLI

Author(s)

AHMADZADEH V. | FARAJNIA S. | HOSSEINPOUR FEIZI M.A. | KHAVARINEJAD R.A. | Issue Writer Certificate

Keywords

HUMANIZATIONQ2

SINGLE-CHAIN ANTIBODY VARIABLE FRAGMENTQ2

CD20Q2

Abstract

Background and Objectives: Rituximab is an anti-CD20 chimeric monoclonal antibody widely used for the treatment of malignant B cells lymphoma. However, the immunogenicity of murine-derived monoclonal antibodies and the large size of full length antibodies restrict cancer immunotherapy. Humanized single chain antibodies can be a solution and a promising alternative for application in immunotherapy. The aim of this study was to produce a humanized scFv antibody for a potential use in the diagnosis and treatment of B cell lymphoma.Materials and Methods: We used a CDR grafting based approach to design a humanized scFv gene fragment. The CDRs were grafted onto the closest human frameworks. The designed sequence was expressed in E. coli then purified. The level of expression was analyzed by SDS-PAGE and the reactivity to CD20 expressing cell line was explored by immunoblotting.Results: Similarity analyses revealed that human germline gene IGHV1-46*03 and IGKV1-39*01 have the highest homology with their murine counterparts. Analysis by SDS-PAGE exhibited a high expression level in E. coli. Reactivity to CD20 expressing Raji cells showed that the produced antibody maintained the binding capacity to human CD20 marker.Conclusion: In our study, humanized anti- CD20 scFv indicated an original antigen-binding affinity. The findings serve as a basis for the development of novel therapeutic strategies in the treatment of CD20- expressing cancers.

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APA: Copy

AHMADZADEH, V., FARAJNIA, S., HOSSEINPOUR FEIZI, M.A., & KHAVARINEJAD, R.A.. (2014). GENERATION OF HUMANIZED SINGLE CHAIN ANTI-CD20ANTIBODY MARKER IN E. COLI. JOURNAL OF ADVANCES IN MEDICAL AND BIOMEDICAL RESEARCH, 21(88), 12-21. SID. https://sid.ir/paper/61613/en

Vancouver: Copy

AHMADZADEH V., FARAJNIA S., HOSSEINPOUR FEIZI M.A., KHAVARINEJAD R.A.. GENERATION OF HUMANIZED SINGLE CHAIN ANTI-CD20ANTIBODY MARKER IN E. COLI. JOURNAL OF ADVANCES IN MEDICAL AND BIOMEDICAL RESEARCH[Internet]. 2014;21(88):12-21. Available from: https://sid.ir/paper/61613/en

IEEE: Copy

V. AHMADZADEH, S. FARAJNIA, M.A. HOSSEINPOUR FEIZI, and R.A. KHAVARINEJAD, “GENERATION OF HUMANIZED SINGLE CHAIN ANTI-CD20ANTIBODY MARKER IN E. COLI,” JOURNAL OF ADVANCES IN MEDICAL AND BIOMEDICAL RESEARCH, vol. 21, no. 88, pp. 12–21, 2014, [Online]. Available: https://sid.ir/paper/61613/en

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