Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays

Key message: Plant expression platform is the new source of immunoglobulin G (IgG) toward small low-molecular-weight targets. The plant-made monoclonal antibody-based immunoassay exhibits comparable analytical performance with hybridoma antibody. Abstract: Immunoassays for small molecules are effici...

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Main Authors: Kaewta Rattanapisit, Tharita Kitisripanya, Atthaphon Konyanee, Worapol Sae-Foo, Apisit Burapapiruin, Waraporn Putalun, Seiichi Sakamoto, Waranyoo Phoolcharoen, Gorawit Yusakul
Other Authors: Chulalongkorn University
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Published: 2022
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Online Access:https://repository.li.mahidol.ac.th/handle/123456789/75697
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spelling th-mahidol.756972022-08-04T14:57:49Z Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays Kaewta Rattanapisit Tharita Kitisripanya Atthaphon Konyanee Worapol Sae-Foo Apisit Burapapiruin Waraporn Putalun Seiichi Sakamoto Waranyoo Phoolcharoen Gorawit Yusakul Chulalongkorn University Walailak University Khon Kaen University Mahidol University Kyushu University Agricultural and Biological Sciences Key message: Plant expression platform is the new source of immunoglobulin G (IgG) toward small low-molecular-weight targets. The plant-made monoclonal antibody-based immunoassay exhibits comparable analytical performance with hybridoma antibody. Abstract: Immunoassays for small molecules are efficiently applied for monitoring of serum therapeutic drug concentration, food toxins, environmental contamination, etc. Immunoglobulin G (IgG) is usually produced using hybridoma cells, which requires complicated procedures and expensive equipment. Plants can act as alternative and economic hosts for IgG production. However, the production of free hapten (low-molecular-weight target)-recognizing IgG from plants has not been successfully developed yet. The current study aimed at creating a plant platform as an affordable source of IgG for use in immunoassays and diagnostic tools. The functional IgG was expressed in Nicotiana benthamiana leaves infiltrated with Agrobacterium tumefaciens strain GV3101 with recombinant geminiviral vectors (pBY3R) occupying chimeric anti-miroestrol IgG genes. The appropriate assembly between heavy and light chains was achieved, and the yield of expression was 0.57 µg/g fresh N. benthamiana leaves. The binding characteristics of the IgG to miroestrol and binding specificity to related compounds, such as isomiroestrol and deoxymiroestrol, were similar to those of hybridoma-produced IgG (monoclonal antibody, mAb). The plant-based mAbs exhibited high sensitivity for miroestrol (IC50, 23.2 ± 2.1 ng/mL), precision (relative standard deviation ≤ 5.01%), and accuracy (97.8–103% recovery), as determined using quantitative enzyme-linked immunosorbent assay. The validated enzyme-linked immunosorbent assay was applicable to determine miroestrol in plant samples. Overall, the plant-produced functional IgG conserved the binding activity and specificity of the parent IgG derived from mammalian cells. Therefore, the plant expression system may be an efficient and affordable platform for the production of antibodies against low-molecular-weight targets in immunoassays. 2022-08-04T07:57:48Z 2022-08-04T07:57:48Z 2021-04-01 Article Plant Cell Reports. Vol.40, No.4 (2021), 723-733 10.1007/s00299-021-02670-z 1432203X 07217714 2-s2.0-85101029723 https://repository.li.mahidol.ac.th/handle/123456789/75697 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85101029723&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 Agricultural and Biological Sciences
spellingShingle Agricultural and Biological Sciences
Kaewta Rattanapisit
Tharita Kitisripanya
Atthaphon Konyanee
Worapol Sae-Foo
Apisit Burapapiruin
Waraporn Putalun
Seiichi Sakamoto
Waranyoo Phoolcharoen
Gorawit Yusakul
Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
description Key message: Plant expression platform is the new source of immunoglobulin G (IgG) toward small low-molecular-weight targets. The plant-made monoclonal antibody-based immunoassay exhibits comparable analytical performance with hybridoma antibody. Abstract: Immunoassays for small molecules are efficiently applied for monitoring of serum therapeutic drug concentration, food toxins, environmental contamination, etc. Immunoglobulin G (IgG) is usually produced using hybridoma cells, which requires complicated procedures and expensive equipment. Plants can act as alternative and economic hosts for IgG production. However, the production of free hapten (low-molecular-weight target)-recognizing IgG from plants has not been successfully developed yet. The current study aimed at creating a plant platform as an affordable source of IgG for use in immunoassays and diagnostic tools. The functional IgG was expressed in Nicotiana benthamiana leaves infiltrated with Agrobacterium tumefaciens strain GV3101 with recombinant geminiviral vectors (pBY3R) occupying chimeric anti-miroestrol IgG genes. The appropriate assembly between heavy and light chains was achieved, and the yield of expression was 0.57 µg/g fresh N. benthamiana leaves. The binding characteristics of the IgG to miroestrol and binding specificity to related compounds, such as isomiroestrol and deoxymiroestrol, were similar to those of hybridoma-produced IgG (monoclonal antibody, mAb). The plant-based mAbs exhibited high sensitivity for miroestrol (IC50, 23.2 ± 2.1 ng/mL), precision (relative standard deviation ≤ 5.01%), and accuracy (97.8–103% recovery), as determined using quantitative enzyme-linked immunosorbent assay. The validated enzyme-linked immunosorbent assay was applicable to determine miroestrol in plant samples. Overall, the plant-produced functional IgG conserved the binding activity and specificity of the parent IgG derived from mammalian cells. Therefore, the plant expression system may be an efficient and affordable platform for the production of antibodies against low-molecular-weight targets in immunoassays.
author2 Chulalongkorn University
author_facet Chulalongkorn University
Kaewta Rattanapisit
Tharita Kitisripanya
Atthaphon Konyanee
Worapol Sae-Foo
Apisit Burapapiruin
Waraporn Putalun
Seiichi Sakamoto
Waranyoo Phoolcharoen
Gorawit Yusakul
format Article
author Kaewta Rattanapisit
Tharita Kitisripanya
Atthaphon Konyanee
Worapol Sae-Foo
Apisit Burapapiruin
Waraporn Putalun
Seiichi Sakamoto
Waranyoo Phoolcharoen
Gorawit Yusakul
author_sort Kaewta Rattanapisit
title Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
title_short Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
title_full Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
title_fullStr Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
title_full_unstemmed Plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin G against small-molecule targets in immunoassays
title_sort plant-made antibody against miroestrol: a new platform for expression of full-length immunoglobulin g against small-molecule targets in immunoassays
publishDate 2022
url https://repository.li.mahidol.ac.th/handle/123456789/75697
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