Abstract
Prostate-specific antigen (PSA) is a protein highly expressed in cancer cells of the prostate gland. In this study, the PSA recombinant protein was fused to the human immunoglobulin G crystallizable fragment (Fc) with the endoplasmic reticulum (ER) Lys-Asp-Glu-Leu (KDEL) retention signal tag for the plant expression system. Agrobacterium-mediated plant transformation was applied to generate transgenic tobacco plants expressing PSA tagged to KDEL (PSAK), PSA-Fc, and PSA-Fc fused to KDEL (PSA-FcK). Western blot analysis showed that the PSA-FcK protein was the most highly expressed among the three different proteins in the transgenic plant leaf. The PSAK, PSA-Fc, and PSA- FcK proteins were successfully purified from plant leaves. Reduced and non-reduced sodium dodecyl sulfate–polyacrylamide gel electrophoresis revealed that the PSA proteins fused to Fc and FcK were dimerized, whereas the PSA itself was not dimerized. N-glycan analysis demonstrated that the N-glycan profile of the PSA-Fc was mainly the structure with α(1,3)-fucose (Fuc) or β(1,2)-xylose (Xyl) glycan residues, whereas the PSA-Fc tagged with the KDEL ER retention signal had mainly an oligomannose-type structure. ELISA and SPR showed that PSA-FcK had a higher binding activity to Fc gamma receptor I than PSA-Fc and human Fc. In addition, PSA-FcK induced anti-PSA, and thus has the potential to be utilized as a prostate cancer vaccine.
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The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Funding
This work was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ0162662022)” Korean Rural Development Administration, and the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) [2021R1F1A1063869].
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Conceptualization, CHS, and KK; methodology, KBK, SRP, YJK, DS., and YJS; validation, CHS, KKB, SRP, YKS, and YJK; formal analysis, KBK.; investigation, CHS., YKS and DSK.; resources, KBK, YKL, KMK, and KK.; data curation, KBK, and YJK; writing—original draft preparation, CHS; writing—review and editing, KSK.; visualization, KBK., and KMK; supervision, KSK; project administration, KBK., and YJK.
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Shin, C., Kim, K., Kang, Y.J. et al. Effect of IgG Fc-fusion and KDEL-ER retention signal on prostate-specific antigen expression in plant and its immune in mice. Plant Biotechnol Rep 16, 729–740 (2022). https://doi.org/10.1007/s11816-022-00810-9
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DOI: https://doi.org/10.1007/s11816-022-00810-9