Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS

Anti-PD-1 and anti-PD-L1 antibodies have achieved great clinical success in cancer immunotherapy, and peptide and small molecule inhibitors of PD-1/PD-L1 binding also attract much attention. Ascidians are not only seafood, but are also an important source of bioactive substances, including anti-tumo...

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Main Authors: Qiuyang Huang, Xiaoling Zang, Xinyu Jin, Qian Liu, Xin Zhang, Xinyu Li, Lizhen Zhao, Zhihua Lv
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Marine Drugs
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Online Access:https://www.mdpi.com/1660-3397/23/4/137
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author Qiuyang Huang
Xiaoling Zang
Xinyu Jin
Qian Liu
Xin Zhang
Xinyu Li
Lizhen Zhao
Zhihua Lv
author_facet Qiuyang Huang
Xiaoling Zang
Xinyu Jin
Qian Liu
Xin Zhang
Xinyu Li
Lizhen Zhao
Zhihua Lv
author_sort Qiuyang Huang
collection DOAJ
description Anti-PD-1 and anti-PD-L1 antibodies have achieved great clinical success in cancer immunotherapy, and peptide and small molecule inhibitors of PD-1/PD-L1 binding also attract much attention. Ascidians are not only seafood, but are also an important source of bioactive substances, including anti-tumor components. In this study, ascidian enzymatic hydrolysates were found to contain PD-1/PD-L1 inhibitory components. Affinity ultrafiltration (AUF) coupled with the nanoLC-MS/MS method was first applied in screening for PD-L1 peptide inhibitors from ascidian enzymatic hydrolysates. Two anti-PD-L1 ascidian peptides, C5 (LDVVIHTVTYGDR) and S2 (VLRDNIQGITKPAIR), were filtered out from the ascidians <i>Ciona intestinalis</i> and <i>Styela clava</i>, respectively. C5 and S2 showed moderate anti-PD-1/PD-L1 effects with the IC<sub>50</sub> values of 33.9 µM (C5) and 112.8 μM (S2), respectively, by homogenous time-resolved fluorescence (HTRF) binding assay, and the K<sub>D</sub> values of 22.9 µM (C5) and 29.1 µM (S2), respectively, by surface plasmon resonance (SPR) assay. The results of this study suggest that ascidian enzymatic hydrolysates may be a potential source of bioactive peptides with anti-PD-1/PD-L1 activity.
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publishDate 2025-03-01
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spelling doaj-art-4136a1e6bbb949f8a65e48ee0b1e5bcc2025-08-20T02:18:09ZengMDPI AGMarine Drugs1660-33972025-03-0123413710.3390/md23040137Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MSQiuyang Huang0Xiaoling Zang1Xinyu Jin2Qian Liu3Xin Zhang4Xinyu Li5Lizhen Zhao6Zhihua Lv7School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaCollege of Physics, Qingdao University, Qingdao 266071, ChinaSchool of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, ChinaAnti-PD-1 and anti-PD-L1 antibodies have achieved great clinical success in cancer immunotherapy, and peptide and small molecule inhibitors of PD-1/PD-L1 binding also attract much attention. Ascidians are not only seafood, but are also an important source of bioactive substances, including anti-tumor components. In this study, ascidian enzymatic hydrolysates were found to contain PD-1/PD-L1 inhibitory components. Affinity ultrafiltration (AUF) coupled with the nanoLC-MS/MS method was first applied in screening for PD-L1 peptide inhibitors from ascidian enzymatic hydrolysates. Two anti-PD-L1 ascidian peptides, C5 (LDVVIHTVTYGDR) and S2 (VLRDNIQGITKPAIR), were filtered out from the ascidians <i>Ciona intestinalis</i> and <i>Styela clava</i>, respectively. C5 and S2 showed moderate anti-PD-1/PD-L1 effects with the IC<sub>50</sub> values of 33.9 µM (C5) and 112.8 μM (S2), respectively, by homogenous time-resolved fluorescence (HTRF) binding assay, and the K<sub>D</sub> values of 22.9 µM (C5) and 29.1 µM (S2), respectively, by surface plasmon resonance (SPR) assay. The results of this study suggest that ascidian enzymatic hydrolysates may be a potential source of bioactive peptides with anti-PD-1/PD-L1 activity.https://www.mdpi.com/1660-3397/23/4/137<i>Ciona intestinalis</i><i>Styela clava</i>PD-L1 peptide inhibitorsaffinity ultrafiltration (AUF)-nanoLC-MS/MShydrolysatestrypsin
spellingShingle Qiuyang Huang
Xiaoling Zang
Xinyu Jin
Qian Liu
Xin Zhang
Xinyu Li
Lizhen Zhao
Zhihua Lv
Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
Marine Drugs
<i>Ciona intestinalis</i>
<i>Styela clava</i>
PD-L1 peptide inhibitors
affinity ultrafiltration (AUF)-nanoLC-MS/MS
hydrolysates
trypsin
title Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
title_full Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
title_fullStr Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
title_full_unstemmed Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
title_short Discovery of PD-L1 Peptide Inhibitors from Ascidian Enzymatic Hydrolysates by Affinity Ultrafiltration Coupled to NanoLC-MS/MS
title_sort discovery of pd l1 peptide inhibitors from ascidian enzymatic hydrolysates by affinity ultrafiltration coupled to nanolc ms ms
topic <i>Ciona intestinalis</i>
<i>Styela clava</i>
PD-L1 peptide inhibitors
affinity ultrafiltration (AUF)-nanoLC-MS/MS
hydrolysates
trypsin
url https://www.mdpi.com/1660-3397/23/4/137
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