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Chimeric single-chain variable fragment-human immunoglobulin G crystallizable fragment antibody against GD2 for neuroblastoma targeted immunotherapy
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Document Title
Chimeric single-chain variable fragment-human immunoglobulin G crystallizable fragment antibody against GD2 for neuroblastoma targeted immunotherapy
Author
Laopajon W. Takheaw N. Kotemul K. Pata S. Hongeng S. Kasinrerk W.
Affiliations
Division of Clinical Immunology Department of Medical Technology Faculty of Associated Medical Sciences Chiang Mai University Chiang Mai 50200 Thailand; Biomedical Technology Research Center National Center for Genetic Engineering and Biotechnology National Science and Technology Development Agency Faculty of Associated Medical Sciences Chiang Mai University Chiang Mai 50200 Thailand; Department of Pediatrics Faculty of Medicine Ramathibodi Hospital Mahidol University Bangkok 10400 Thailand
Type
Article
Source Title
Exploration of Targeted Anti-tumor Therapy
ISSN
26923114
Year
2023
Volume
4
Issue
5
Page
1145-1156
Open Access
All Open Access Gold Green
Publisher
Open Exploration Publishing Inc
DOI
10.37349/etat.2023.00188
Abstract
Aim: The present study aims to generate chimeric mouse single-chain variable fragment (scFv) and immunoglobulin G1 (IgG1) crystallizable fragment (Fc) antibody against disialoganglioside (GD2) for the treatment of neuroblastoma (NB). The generated scFv-IgG Fc antibody lacking first constant domain of heavy chain (CH1) is of a smaller size than the natural antibody and has anti-tumor activity. Methods: Vector for scFv-IgG Fc antibody was constructed and scFv-IgG Fc antibody was expressed in human embryonic kidney 293T (HEK293T) cell line. Purification of scFv-IgG Fc antibody from the culture supernatant of transfected HEK293T cells was performed by Protein G affinity chromatography. The structure and binding activity of scFv-IgG Fc antibody were verified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) western blotting (WB) and immunofluorescence techniques. Anti-tumor activities by antibody-dependent cellular cytotoxicity (ADCC) and antibodydependent cellular phagocytosis (ADCP) were determined. Results: Using plasmid fusion-human IgG1-Fc2 tag vector (pFUSE-hIgG1-Fc2) a plasmid vector encoding chimeric mouse scFv and hIgG1 Fc antibody against GD2 was successfully constructed. This vector was transfected into human HEK293T cells to produce scFv-IgG Fc antibody. The transfected HEK293T cells could produce chimeric scFv-IgG Fc antibody against GD2 which lacks the IgG heavy chain CH1 domain but carries CH2 and CH3 domains. The chimeric antibodies could be purified from the culture supernatant of the transfected HEK293T culture in the presence of zeocin drug. The produced GD2 scFv-IgG Fc antibodies which are smaller in size than the intact antibody could trigger the killing of GD2 expressed NB cell line SHSY5Y by ADCC and ADCP mechanisms. Conclusions: The results indicate that chimeric scFv-hIgG Fc antibody lacking heavy chain CH1 domain could mediate antibody induced anti-tumor activities. The small size of this type of chimeric antibody may be employed as anti-GD2 antibody for NB therapy. ? The Author(s) 2023.
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
License
CC BY
Rights
Authors
Publication Source
Scopus