Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation

In this study, a key issue to be addressed is the safe disposal of hybridoma instability. Hybridoma technology was used to produce anti-O. viverrini monoclonal antibody. Previous studies have shown that antibody production via antibody phage display can sustain the hybridoma technique. This paper pr...

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Main Authors: Sitthinon Siripanthong, Anchalee Techasen, Chanin Nantasenamat, Aijaz Ahmad Malik, Paiboon Sithithaworn, Chanvit Leelayuwat, Amonrat Jumnainsong
Other Authors: Faculty of Medicine, Khon Kaen University
Format: Article
Published: 2022
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/79376
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spelling th-mahidol.793762022-08-04T18:41:23Z Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation Sitthinon Siripanthong Anchalee Techasen Chanin Nantasenamat Aijaz Ahmad Malik Paiboon Sithithaworn Chanvit Leelayuwat Amonrat Jumnainsong Faculty of Medicine, Khon Kaen University Khon Kaen University Mahidol University Multidisciplinary In this study, a key issue to be addressed is the safe disposal of hybridoma instability. Hybridoma technology was used to produce anti-O. viverrini monoclonal antibody. Previous studies have shown that antibody production via antibody phage display can sustain the hybridoma technique. This paper presents the utility of antibody phage display technology for producing the phage displayed KKU505 Fab fragment and using experiments in concomitant with molecular simulation for characterization. The phage displayed KKU505 Fab fragment and characterization were successfully carried out. The KKU505 hybridoma cell line producing anti-O. viverrini antibody predicted to bind to myosin was used to synthesize cDNA so as to amplify the heavy chain and the light chain sequences. The KKU505 displayed phage was constructed and characterized by a molecular modeling in which the KKU505 Fab fragment and -O. viverrini myosin head were docked computationally and it is assumed that the Fab fragment was specific to -O. viverrini on the basis of mass spectrometry and Western blot. This complex interaction was confirmed by molecular simulation. Furthermore, the KKU505 displayed phage was validated using indirect enzyme-linked immunosorbent assays (ELISA) and immunohistochemistry. It is worthy to note that ELISA and immunohistochemistry results confirmed that the Fab fragment was specific to the -O. viverrini antigen. Results indicated that the approach presented herein can generate anti-O. viverrini antibody via the phage display technology. This study integrates the use of phage display technology together with molecular simulation for further development of monoclonal antibody production. Furthermore, the presented work has profound implications for antibody production, particularly by solving the problem of hybridoma stability issues. 2022-08-04T11:41:23Z 2022-08-04T11:41:23Z 2021-03-01 Article PLoS ONE. Vol.16, No.3 March (2021) 10.1371/journal.pone.0248887 19326203 2-s2.0-85103185589 https://repository.li.mahidol.ac.th/handle/123456789/79376 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85103185589&origin=inward
institution Mahidol University
building Mahidol University Library
continent Asia
country Thailand
Thailand
content_provider Mahidol University Library
collection Mahidol University Institutional Repository
topic Multidisciplinary
spellingShingle Multidisciplinary
Sitthinon Siripanthong
Anchalee Techasen
Chanin Nantasenamat
Aijaz Ahmad Malik
Paiboon Sithithaworn
Chanvit Leelayuwat
Amonrat Jumnainsong
Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
description In this study, a key issue to be addressed is the safe disposal of hybridoma instability. Hybridoma technology was used to produce anti-O. viverrini monoclonal antibody. Previous studies have shown that antibody production via antibody phage display can sustain the hybridoma technique. This paper presents the utility of antibody phage display technology for producing the phage displayed KKU505 Fab fragment and using experiments in concomitant with molecular simulation for characterization. The phage displayed KKU505 Fab fragment and characterization were successfully carried out. The KKU505 hybridoma cell line producing anti-O. viverrini antibody predicted to bind to myosin was used to synthesize cDNA so as to amplify the heavy chain and the light chain sequences. The KKU505 displayed phage was constructed and characterized by a molecular modeling in which the KKU505 Fab fragment and -O. viverrini myosin head were docked computationally and it is assumed that the Fab fragment was specific to -O. viverrini on the basis of mass spectrometry and Western blot. This complex interaction was confirmed by molecular simulation. Furthermore, the KKU505 displayed phage was validated using indirect enzyme-linked immunosorbent assays (ELISA) and immunohistochemistry. It is worthy to note that ELISA and immunohistochemistry results confirmed that the Fab fragment was specific to the -O. viverrini antigen. Results indicated that the approach presented herein can generate anti-O. viverrini antibody via the phage display technology. This study integrates the use of phage display technology together with molecular simulation for further development of monoclonal antibody production. Furthermore, the presented work has profound implications for antibody production, particularly by solving the problem of hybridoma stability issues.
author2 Faculty of Medicine, Khon Kaen University
author_facet Faculty of Medicine, Khon Kaen University
Sitthinon Siripanthong
Anchalee Techasen
Chanin Nantasenamat
Aijaz Ahmad Malik
Paiboon Sithithaworn
Chanvit Leelayuwat
Amonrat Jumnainsong
format Article
author Sitthinon Siripanthong
Anchalee Techasen
Chanin Nantasenamat
Aijaz Ahmad Malik
Paiboon Sithithaworn
Chanvit Leelayuwat
Amonrat Jumnainsong
author_sort Sitthinon Siripanthong
title Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
title_short Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
title_full Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
title_fullStr Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
title_full_unstemmed Production and characterization of antibody against Opisthorchis viverrini via phage display and molecular simulation
title_sort production and characterization of antibody against opisthorchis viverrini via phage display and molecular simulation
publishDate 2022
url https://repository.li.mahidol.ac.th/handle/123456789/79376
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