Abstract
Plants can be used to produce recombinant functional proteins such as antibodies and vaccines. In this study, transgenic plants were utilized to express anti-human programmed death ligand-1 (PD-L1) monoclonal antibodies (mAbP PD-L1) tagged with the endoplasmic reticulum retention motif KDEL. PCR and immunoblot analyses confirmed that mAb PD-L1 was expressed in transgenic plants. ELISA analysis confirmed the binding activity of mAbP PD-L1 to human prostate and urothelial cancer cell lines PC-3 and TCCSUP, respectively. Confocal imaging analysis demonstrated that mAbP PD-L1 had binding activity against human cancer cell lines similar to Avelumab (mAbM PD-L1). Confocal imaging analysis showed that the binding activity of mAbP PD-L1 to the human tonsil is comparable to mAbM PD-L1. These results suggest that mAbP PD-L1 shows functional binding activities for PD-L1+ cancer cells. Hence, this study showed that tobacco plants can be used to produce functional anti-human PD-L1 mAbs.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benicky J, Sanda M, Brnakova Kennedy Z et al (2020) PD-L1 glycosylation and its impact on binding to clinical antibodies. J Proteome Res 20:485–497
Boyerinas B, Jochems C, Fantini M et al (2015) Antibody-dependent cellular cytotoxicity activity of a novel anti-PD-L1 antibody avelumab (MSB0010718C) on human tumor cells an anti-PD-L1 mAb that mediates ADCC of human tumor cells. Cancer Immunol Res 3:1148–1157
Chen C-L, Pan Q-Z, Zhao J-J et al (2016) PD-L1 expression as a predictive biomarker for cytokine-induced killer cell immunotherapy in patients with hepatocellular carcinoma. Oncoimmunology 5:e1176653
Chung HJ, Noh Y, Kim MS et al (2018) Steroidogenic effects of taraxacum officinale extract on the levels of steroidogenic enzymes in mouse Leydig cells. Anim Cells Syst 22:407–414
Datla RS, Bekkaoui F, Hammerlindl JK, Pilate G, Dunstan DI, Crosby WLJ (1993) Improved high-level constitutive foreign gene expression in plants using an AMV RNA4 untranslated leader sequence. Plant Sci 94:139–149
Escors D, Gato-Cañas M, Zuazo M et al (2018) The intracellular signalosome of PD-L1 in cancer cells. Signal Transd Target Ther 3:1–9
Grabowski GA, Golembo M, Shaaltiel YJ (2014) Taliglucerase alfa: an enzyme replacement therapy using plant cell expression technology. Mol Genet Metab 112:1–8
He J, Hu Y, Hu M, Li BJ (2015) Development of PD-1/PD-L1 pathway in tumor immune microenvironment and treatment for non-small cell lung cancer. Sci Rep 5:1–9
Janse Van Rensburg HJ, Azad T, Ling M et al (2018) The hippo pathway component TAZ promotes immune evasion in human cancer through PD-L1TAZ regulates cancer immune evasion through PD-L1. Cancer Res 78:1457–1470
Jiang X, Liu G, Li Y, Pan YJG (2021) Immune checkpoint: the novel target for antitumor therapy. Genes Dis 8:25–37
Keilholz U, Mehnert JM, Bauer S et al (2019) Avelumab in patients with previously treated metastatic melanoma: phase 1b results from the JAVELIN Solid Tumor trial. J Immunother Cancer 7:1–13
Ko K (2014) Expression of recombinant vaccines and antibodies in plants. Monoclonal Antibodies Immunodiagn Immunother 33:192–198
Ko K, Koprowski HJVR (2005) Plant biopharming of monoclonal antibodies. Virus Res 111:93–100
Lee JH, Ko KJB, Therapeutics, (2017) Production of recombinant anti-cancer vaccines in plants. Biomol Ther 25:345
Lee J-H, Park D-Y, Lee K-J et al (2013) Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody. PLoS ONE 8:e68772
Lee J, Han JH, Jang A, Kim JW, Hong SA, Myung SCJPO (2016) DNA methylation-mediated downregulation of DEFB1 in prostate cancer cells. PLoS ONE 11:e0166664
Lim C-Y, Kim D-S, Kang Y et al (2022) Immune responses to plant-derived recombinant colorectal cancer glycoprotein EpCAM-FcK fusion protein in mice. Biomol Ther 30:546–552
Miao J, Hsu P-C, Yang Y-L et al (2017) YAP regulates PD-L1 expression in human NSCLC cells. Oncotarget 8:114576
Park S-R, Lim C-Y, Kim D-S, Ko KJFIPS (2015) Optimization of ammonium sulfate concentration for purification of colorectal cancer vaccine candidate recombinant protein GA733-FcK isolated from plants. Front Plant Sci 6:1040
Rademacher T, Sack M, Blessing D, Fischer R, Holland T, Buyel J (2019) Plant cell packs: a scalable platform for recombinant protein production and metabolic engineering. Plant Biotechnol J 17:1560–1566
Sasidharan Nair V, Elkord EJI, Biology C (2018) Immune checkpoint inhibitors in cancer therapy: a focus on T-regulatory cells. Immunol Cell Biol 96:21–33
Schillberg S, Finnern RJ (2021) Plant molecular farming for the production of valuable proteins—critical evaluation of achievements and future challenges. J Plant Physiol 258:153359
So Y, Lee K-J, Kim D-S et al (2013) Glycomodification and characterization of anti-colorectal cancer immunotherapeutic monoclonal antibodies in transgenic tobacco. Plant Cell Tissue Organ Culture 113:41–49
Temporini C, Colombo R, Calleri E et al (2020) Chromatographic tools for plant-derived recombinant antibodies purification and characterization. J Pharm Biomed Anal 179:112920
Verma R, Boleti E, George AJ (1998) Antibody engineering: comparison of bacterial, yeast, insect and mammalian expression systems. J Immunol Methods 216:165–181
Funding
This work was supported by a National Research Foundation of Korea grant funded by the Korean Government (MEST) (NRF-2020R1A2C2010652) and the Chung-Ang University Research Scholarship Grants in 2020.
Author information
Authors and Affiliations
Contributions
Conceptualization, CEL, KK, and SCM; methodology, KBK, JHL, HJC, DWL, JSL, KSK, HJM, and CEL; validation, JWK, HJC, and YSL; formal analysis, JHL, and CEL; investigation, KK, and SCM; resources, HJM, KK, and SCM; data curation, CEL, and KBK; writing—original draft preparation, CEL; writing—review and editing, SCM; and KK; visualization, CEL; supervision, SCM; project administration, SCM.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Lee, C.E., Lee, J.H., Chung, H.J. et al. Expression and in vitro function of anti-PD-L1 human antibody expressed in plant. Plant Biotechnol Rep 17, 531–539 (2023). https://doi.org/10.1007/s11816-023-00844-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11816-023-00844-7