Journal of Peptide Science最新文献

N- to C-Peptide Synthesis, Arguably the Future for Sustainable Production N- to - c肽合成，可持续生产的未来

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-05-01 DOI: 10.1002/psc.70019

Kinshuk Ghosh, William D. Lubell

{"title":"N- to C-Peptide Synthesis, Arguably the Future for Sustainable Production","authors":"Kinshuk Ghosh, William D. Lubell","doi":"10.1002/psc.70019","DOIUrl":"https://doi.org/10.1002/psc.70019","url":null,"abstract":"A revolution in peptide production arrived from the innovation of carboxylate to amine C- to N-direction solid-phase synthesis. This cornerstone of modern peptide science has enabled multiple academic and industrial applications; however, the process of C- to N-solid phase peptide synthesis (C-N-SPPS) has extreme process mass intensity and poor atom economy. Notably, C-N-SPPS relies upon the use of atom-intensive protecting groups, such as the fluorenylmethyloxycarbonyl (Fmoc) protection and wasteful excess of protected amino acids and coupling agents. On the other hand, peptide synthesis in the amine to carboxylate N- to C-direction offers potential to minimize protection and may arguably enable more efficient means for manufacturing peptides. For example, efficient amide bond formation in the N- to C-direction has been accomplished using methods employing thioesters, vinyl esters, and transamidation to achieve peptide synthesis with minimal epimerization. This review aims to provide an overview of N- to C-peptide synthesis indicating advantages in taking this avenue for sustainable peptide production.","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/psc.70019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143897104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Antimicrobial Activity of Short Analogues of the Marine Peptide EeCentrocin 1: Synthesis of Lipopeptides and Head-to-Tail Cyclic Peptides and Mechanism of Action Studies 海洋肽eecentrrocin 1短类似物的抗菌活性：脂肽和首尾环肽的合成及其作用机制研究

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-28 DOI: 10.1002/psc.70025

Danijela Simonovic, Hymonti Dey, Natascha Johansen, Trude Anderssen, Ida K. Ø. Hansen, Hege Devold, Terje Vasskog, Hans-Matti Blencke, Frode Jacobsen Øyen, Elizabeth G. Aarag Fredheim, Tor Haug, Morten B. Strøm

{"title":"Antimicrobial Activity of Short Analogues of the Marine Peptide EeCentrocin 1: Synthesis of Lipopeptides and Head-to-Tail Cyclic Peptides and Mechanism of Action Studies","authors":"Danijela Simonovic, Hymonti Dey, Natascha Johansen, Trude Anderssen, Ida K. Ø. Hansen, Hege Devold, Terje Vasskog, Hans-Matti Blencke, Frode Jacobsen Øyen, Elizabeth G. Aarag Fredheim, Tor Haug, Morten B. Strøm","doi":"10.1002/psc.70025","DOIUrl":"https://doi.org/10.1002/psc.70025","url":null,"abstract":"<div>\u0000 \u0000 We have synthesised a series of 12-residue analogues of a previously reported lead peptide (P6) developed from the heavy chain of the marine peptide EeCentrocin 1, isolated from the sea urchin Echinus esculentus. We optimised the lead peptide by increasing its net positive charge, its lipophilicity through N-terminal fatty acid acylation or incorporation of a Trp residue, and by synthesising head-to-tail cyclic peptides under pseudo–high-dilution conditions. All peptides were screened for antimicrobial and antifungal activity, and toxicity was determined against human red blood cells. The two most potent peptide analogues were the linear peptide P6-W6R8 and its head-to-tail cyclic analogue cP6-W6R8 displaying minimum inhibitory concentrations of 0.4–6.6 μM against Gram-positive and Gram-negative bacteria and 6.2–13 μM against fungi. All peptides showed low haemolytic activity except for two of the lipopeptides, in which haemolytic activity correlated with increasing acyl chain length. Mode of action studies using bacterial biosensor strains revealed a membrane disruptive effect of both the linear and the cyclic peptide. Overall, the results of our study demonstrated that relatively simple structural modifications could be successfully employed in the development of potent antimicrobial lead peptides derived from marine natural products.\u0000 </div>","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143883965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Efficient Screening of α-Glucosidase Inhibitory Peptides From Seahorse Through the Innovative Joint Technique: De Novo Sequencing and Parallel SPOT Synthesis 通过创新的联合技术：从头测序和平行点合成高效筛选海马α-葡萄糖苷酶抑制肽

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-28 DOI: 10.1002/psc.70023

Shengfang Gao, Yimeng Li, Xiaohui Zhang, Zhuo Cao, Youyou Guo, Runkun Zhao, Lifan Li, Hongying Lin, Qi Qin, Bingqing Yi, Guodong Zhao

{"title":"Efficient Screening of α-Glucosidase Inhibitory Peptides From Seahorse Through the Innovative Joint Technique: De Novo Sequencing and Parallel SPOT Synthesis","authors":"Shengfang Gao, Yimeng Li, Xiaohui Zhang, Zhuo Cao, Youyou Guo, Runkun Zhao, Lifan Li, Hongying Lin, Qi Qin, Bingqing Yi, Guodong Zhao","doi":"10.1002/psc.70023","DOIUrl":"https://doi.org/10.1002/psc.70023","url":null,"abstract":"<div>\u0000 \u0000 In this research, de novo sequencing was innovatively combined with parallel SPOT synthesis for the efficient screening of biological peptides from TCM or seafood: seahorse with synergistic antioxidant and α-glucosidase inhibitory activities, which is promising for postprandial hyperglycemia management. Gastrointestinal digestion mimic and de novo sequencing were sequentially carried out to predict new peptides from seahorse. After bioinformatic analysis using Peptide Ranker, 82 peptides were eventually synthesized by efficient parallel SPOT technique, and Ser-Val-Try-Leu-Gly-Gly-Ser-Leu-Leu (SVWLGGSLL) was screened out as the most efficient peptide with synergistic antioxidant (DPPH radical scavenging activity of 77%) and α-glucosidase inhibitory activity (IC50 = 0.36 mM). Molecular docking was further carried out to illustrate the favorable ligand-receptor interactions formed such as hydrogen bonding and van der Waals force with low binding free energy of −7.8 kcal/mol. Moreover, pharmacokinetic analysis indicated that SVWLGGSLL was unrelated to toxicity with the advantage of gastrointestinal stability.\u0000 </div>","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143880150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Helicity-Dependent Enzymatic Peptide Cyclization 螺旋依赖性酶促肽环化

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-27 DOI: 10.1002/psc.70024

Canan Durukan, Jannik Faierson, Isabel van der Wal, Juan Lizandra Pérez, Sven Hennig, Tom N. Grossmann

引用次数: 0

Peptide-Based Strategies in PLGA-Enhanced Tumor Therapy 基于多肽的plga增强肿瘤治疗策略

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-23 DOI: 10.1002/psc.70020

Hong-Lin Han, Jing-Yun Su, Xiao-Huan Zhao, Dan-Dan Hou, Yan-Mei Li

{"title":"Peptide-Based Strategies in PLGA-Enhanced Tumor Therapy","authors":"Hong-Lin Han, Jing-Yun Su, Xiao-Huan Zhao, Dan-Dan Hou, Yan-Mei Li","doi":"10.1002/psc.70020","DOIUrl":"https://doi.org/10.1002/psc.70020","url":null,"abstract":"<div>\u0000 \u0000 Peptide-based therapeutics have gained attention in cancer treatment because of their good specificity, low toxicity, and ability to modulate immune responses. However, challenges such as enzymatic degradation and poor bioavailability limit their clinical application. Peptide-functionalized poly(lactic-co-glycolic acid) (PLGA) systems have emerged as a transformative platform in cancer therapy that offers unique advantages, including enhanced stability, sustained release, and precise delivery of therapeutic agents. This review highlights the synergistic integration of peptides with PLGA and addresses key challenges of peptide-based therapeutics. The application of peptide-functionalized PLGA systems encompasses a diverse range of strategies for cancer therapy. In chemotherapy, peptides disrupt critical tumor pathways, induce apoptosis, and inhibit angiogenesis, demonstrating their versatility in targeting various aspects of tumor progression. In immunotherapy, peptides act as antigens to stimulate robust immune responses or as immune checkpoint inhibitors to restore T cell activity, overcoming tumor immune evasion. These systems also harness the enhanced permeability and retention effect, facilitating preferential accumulation in tumor tissues while leveraging tumor microenvironment (TME)-responsive mechanisms, such as pH-sensitive or enzyme-triggered drug release, to achieve controlled, localized delivery. Collectively, peptide-functionalized PLGA systems represent a promising, versatile approach for precise cancer therapy that integrates innovative delivery strategies with highly specific, potent therapeutic agents.\u0000 </div>","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143865791","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Beyond Efficacy: Ensuring Safety in Peptide Therapeutics through Immunogenicity Assessment 超越功效：通过免疫原性评估确保多肽治疗的安全性

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-21 DOI: 10.1002/psc.70016

Koulla Achilleos, Christos Petrou, Vicky Nicolaidou, Yiannis Sarigiannis

{"title":"Beyond Efficacy: Ensuring Safety in Peptide Therapeutics through Immunogenicity Assessment","authors":"Koulla Achilleos, Christos Petrou, Vicky Nicolaidou, Yiannis Sarigiannis","doi":"10.1002/psc.70016","DOIUrl":"https://doi.org/10.1002/psc.70016","url":null,"abstract":"Peptides are gaining remarkable popularity in clinical diagnosis and treatment due to their high selectivity and minimal side effects. Over 11% of all new pharmaceutical chemical entities authorised by the FDA between 2016 and 2024 were synthetically manufactured peptides. A critical factor that can potentially limit the efficacy and safety of peptide-based therapeutics or biologics is immunogenicity, defined as an unintended or adverse immune response to a protein or peptide therapy. This response may be triggered by the peptide itself or by impurities in the production or formulation steps, leading to the production of antidrug antibodies (ADAs). To address this, current regulatory guidelines require the assessment of risks in market authorization applications, which include identifying drug impurity levels and immunogenicity. The development and critical evaluation of appropriate immunogenicity assays is therefore highly warranted. Such assays must consider the fine complexities of the immune response, as well as its variation within the human population. Moreover, immunogenicity testing is expected to remain a priority as the shift toward greener chemistries in peptide synthesis may require reassessment of novel impurities in peptide formulations.","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/psc.70016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143852715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cyclization of the Analgesic α-Conotoxin Vc1.1 With a Non-Natural Linker: Effects on Structure, Stability, and Bioactivity 非天然连接体环化镇痛药α-蛇毒素Vc1.1：对结构、稳定性和生物活性的影响

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-21 DOI: 10.1002/psc.70017

Yuhui Zhang, Han Shen Tae, David J. Adams, Thomas Durek, David J. Craik

{"title":"Cyclization of the Analgesic α-Conotoxin Vc1.1 With a Non-Natural Linker: Effects on Structure, Stability, and Bioactivity","authors":"Yuhui Zhang, Han Shen Tae, David J. Adams, Thomas Durek, David J. Craik","doi":"10.1002/psc.70017","DOIUrl":"https://doi.org/10.1002/psc.70017","url":null,"abstract":"α-Conotoxin Vc1.1 is a disulfide-rich peptide and a promising drug candidate for treating neuropathic and chronic pain. Backbone cyclization was applied to enhance its drug-like properties, resulting in improved serum stability and oral bioavailability. However, this modification also adversely affected its stability and activity in simulated intestinal fluid (SIF). To address these adverse effects, we explored the use of polyethylene glycol (PEG) linkers as substitutes for peptide backbone cyclization linkers. PEG linkers are smaller, more flexible, and more stable than peptide linkers. Furthermore, previous studies have demonstrated that PEG backbone linkers can enhance the activity of conotoxins. In this study, we synthesized four PEG-backboned cyclic Vc1.1 (cVc1.1) analogues with varying lengths of PEG linkers and used a chemo-enzymatic method to cyclize these analogues. Their structure, stability, and activity were subsequently evaluated. Although the results revealed that PEG linkers preserved the SIF stability and activity of cVc1.1, they highlighted the crucial role of the peptide's helical structure in maintaining its stability and activity. Additionally, this work introduces a novel approach for synthesizing cyclic conotoxins.","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 6","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/psc.70017","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143852714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synthesis and Biological Activity of Ultrashort Antimicrobial Peptides Bearing a Non-Coded Amino Acid 非编码氨基酸超短抗菌肽的合成及其生物活性研究

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-14 DOI: 10.1002/psc.70021

Cristina Peggion, Andrea Schivo, Martina Rotondo, Simona Oancea, Lucia-Florina Popovici, Teodora Călin, Anna Mkrtchyan, Ashot Saghyan, Liana Hayriyan, Emma Khachatryan, Fernando Formaggio, Barbara Biondi

{"title":"Synthesis and Biological Activity of Ultrashort Antimicrobial Peptides Bearing a Non-Coded Amino Acid","authors":"Cristina Peggion, Andrea Schivo, Martina Rotondo, Simona Oancea, Lucia-Florina Popovici, Teodora Călin, Anna Mkrtchyan, Ashot Saghyan, Liana Hayriyan, Emma Khachatryan, Fernando Formaggio, Barbara Biondi","doi":"10.1002/psc.70021","DOIUrl":"https://doi.org/10.1002/psc.70021","url":null,"abstract":"Antimicrobial resistance represents a significant global health threat, prompting the exploration of alternative therapeutic strategies. Antimicrobial peptides (AMPs) and lipopeptides are promising candidates due to their unique ability to disrupt bacterial cell membranes through mechanisms distinct from conventional antibiotics. These peptides are typically enhanced by motifs involving cationic amino acids, positive charge, and aromatic residues. Additionally, the conjugation of acyl chains to the N-terminus of AMPs has been shown to improve their antimicrobial activity and selectivity. However, the susceptibility of peptides to enzymatic degradation presents a major limitation. To address this, we investigated the incorporation of non-coded amino acids (NCAAs) to enhance peptide stability. Specifically, we synthesized the NCAA 2-amino-3-(1H-imidazol-1-yl)propanoic acid [His*], producing both enantiomers with high yield and optical purity. We then designed various analogs of ultra-short AMPs by inserting His* at specific positions, evaluating their antimicrobial properties with different acyl chain lengths (C16 and C12) at the N-terminus and the C-terminus. We were able to identify a very promising candidate for applications (P8) characterized by resistance to proteolysis and enhanced biological effectiveness.","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 5","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/psc.70021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143831308","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Synthesis of a Trans-Phakellistatin 21/22 Conformer and Related Alanine Scanning Analogs With Neuroprotective Activity 具有神经保护作用的反式phakellistatin 21/22构象及相关丙氨酸扫描类似物的合成

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-14 DOI: 10.1002/psc.70018

Iqra Kanwal, Hunain Ali, Farkhanda Mushtaq, Sadia Basharat Ali, A. Ganesan, Shabana Usman Simjee, Muhammad Adnan Akram, Farzana Shaheen

{"title":"Synthesis of a Trans-Phakellistatin 21/22 Conformer and Related Alanine Scanning Analogs With Neuroprotective Activity","authors":"Iqra Kanwal, Hunain Ali, Farkhanda Mushtaq, Sadia Basharat Ali, A. Ganesan, Shabana Usman Simjee, Muhammad Adnan Akram, Farzana Shaheen","doi":"10.1002/psc.70018","DOIUrl":"https://doi.org/10.1002/psc.70018","url":null,"abstract":"<div>\u0000 \u0000 The phakellistatins are a class of cyclic peptide natural products among which phakellistatin 21 and 22 isolated from the marine sponge Stylissa flabelliformis are cyclo(Pro1-Pro2-Met(O)3-Phe4-Glu5-Leu6-Pro7-Pro8-Tyr9-Ile10) epimeric at the methionine sulfoxide residue. The natural product contains two cis and two trans proline residues and is reported to have significant cytotoxic activities. We attempted the total synthesis of phakellistatin 21/22 via on-resin macrocyclization using methionine as a building block. The final product contained methionine sulfoxide, suggesting that aerial oxidation took place during the synthesis and during the original isolation of the natural product. Our synthetic peptide cyclo(trans-Pro1,2,7,8)-Pha21 (1) was identified as an unnatural conformer of natural product phakellistatin 21/22 with all Pro residues present as trans amides. The Peptide 1 was inactive against human cancer cell lines, unlike the natural product. We additionally synthesized alanine scanning Analogs 2–5 in which a Pro residue was replaced by Ala and Analog 6, where all four Pro residues were substituted by Ala. Peptides 1, 2, 3, and 5 were found to have neuroprotective effects on primary cortex cells and are potential leads for the treatment of neurodegenerative disorders.\u0000 </div>","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 5","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143831307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Penta-ALFA-Tagged Substrates for Self-Labelling Tags Allow Signal Enhancement in Microscopy 用于自标记标签的penta - alfa标记底物允许在显微镜下增强信号

IF 1.8 4区生物学

Journal of Peptide Science Pub Date : 2025-04-13 DOI: 10.1002/psc.70015

Souvik Ghosh, Ramona Birke, Ashwin Karthick Natarajan, Johannes Broichhagen

{"title":"Penta-ALFA-Tagged Substrates for Self-Labelling Tags Allow Signal Enhancement in Microscopy","authors":"Souvik Ghosh, Ramona Birke, Ashwin Karthick Natarajan, Johannes Broichhagen","doi":"10.1002/psc.70015","DOIUrl":"https://doi.org/10.1002/psc.70015","url":null,"abstract":"Self-labelling proteins like SNAP- and HaloTag have advanced imaging in life sciences by enabling live-cell labeling with fluorophore-conjugated substrates. However, the typical one-fluorophore-per-protein system limits signal intensity. To address this, we developed a strategy using the ALFA-tag system, a 13-amino acid peptide recognized by a bio-orthogonal and fluorescently labelled nanobody, for signal amplification. We synthesized a pentavalent ALFA5 peptide and used an azidolysine for conjugation with a Cy5-modified SNAP- or HaloTag ligand through strain-promoted click chemistry. In vitro measurements on SDS-PAGE showed labelling, and the peptides covalently reacted with their respective tag. HEK293 cells expressing SNAP- and HaloTag-mGluR2 fusion proteins were labeled with ALFA5-Cy5 substrates, and confocal microscopy revealed a significant enhancement in the far-red signal intensity upon nanobody addition, as quantified by integrated signal density ratios. Comparisons between SNAP- and HaloTag substrates showed superior performance for the latter, achieving better signal-to-noise and signal-to-background ratios, as well as overall signal intensity in plasma membrane-localized regions. Our results demonstrate the potential of ALFA-tag-based systems to amplify SLP fluorescent signals. This strategy combines the photostability of synthetic fluorophores with multivalent labeling, providing a powerful tool for advanced imaging applications including super-resolution in cells. Its versatility is expandable across diverse protein systems and colors.","PeriodicalId":16946,"journal":{"name":"Journal of Peptide Science","volume":"31 5","pages":""},"PeriodicalIF":1.8,"publicationDate":"2025-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/psc.70015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143826887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0