Protein and Peptide Letters最新文献

{"title":"Holographic Proteomics: A Review of Digital Holographic Microscopy Applications in Spatial Proteomics.","authors":"Feng Zhu, Qingwen Wang, Xuejia Zheng, Ruiyuan Chen, Chengcheng Liu, Liu Xiang, Jingquan He, Yong Dai","doi":"10.2174/0109298665404512251021101809","DOIUrl":"10.2174/0109298665404512251021101809","url":null,"abstract":"<p><p>Holoproteomics is a state-of-the-art advancement in spatial proteomics, enabling comprehensive spatial analysis of proteins in tissue microenvironments by combining Digital Holographic Microscopy (DHM) imaging and high-precision laser capture microdissection (LCM) techniques. DHM is an advanced technology that utilizes optical interference principles for non-invasive imaging, providing high-resolution three-dimensional visualization of cells and macromolecules without requiring markers. In proteomics, DHM provides crucial technical support for investigating protein interactions and enables high-precision tracking and analysis of dynamic protein changes. In this review, we systematically survey peer-reviewed literature published in the past five years, with a focus on experimental and clinical studies applying DHM to proteomic analyses. Based on the significant advantages of this technology, we introduce the concept of \"Holographic proteomics\" as an emerging research field with promising future directions.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"693-710"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145524182","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}

{"title":"A Preliminary Study on the Antibacterial Activity of the Secretion of the Levantine Water Frog, <i>Pelophylax bedriagae</i> (Camerano, 1882) (Anura:Ranidae).","authors":"Nazlı Atci, Ebru Tanriverdio, Dincer Ayaz","doi":"10.2174/0109298665385364250917100034","DOIUrl":"10.2174/0109298665385364250917100034","url":null,"abstract":"<p><strong>Introduction: </strong>The skin of amphibians performs some vital roles, such as camouflage, ion and water transport, and gas exchange. Additionally, it plays a significant role in the immune system by preventing pathogen invasion. The secretions produced by the granular glands in the skin possess antimicrobial properties, which help prevent harmful microorganisms from entering the animal's body. The study aims to determine the total protein amounts in the secretion of Pelophylax bedriagae (Levant water frog) distributed in Türkiye and to reveal whether it has antimicrobial properties. In this context, it is a pioneering study on antimicrobial peptides in the skin secretion of Pelophylax.</p><p><strong>Methods: </strong>Field research was carried out at İzmir, Türkiye's Homeros Valley. Because AMPs are species-specific, not sex-specific peptides, both male and female adults were used, regardless of sex. The average weight of the animals used in the experiment was 21 grams. Granular glands were stimulated using a specialized device, and the secretions were collected for analysis. The collected secretions were lyophilized after centrifugation. Then, the total protein amount in the secretion was determined by Bicinchoninic Acid (BCA). The antibacterial activities of the skin secretions against <i>Escherichia coli</i> and <i>Staphylococcus aureus</i> were assessed using a plate well diffusion assay. The peptide profiles in the skin secretions were determined using the Tricine-SDS-- PAGE electrophoresis method.</p><p><strong>Results: </strong>The plate well diffusion assay demonstrated that the skin secretions created a 21 mm inhibition zone against E. coli and a 20 mm inhibition zone against S. aureus. The results of the Tricine-SDS-PAGE electrophoresis revealed the presence of peptides with five different molecular weights, including one smaller than 5 kDa. As a result of the analysis, it was determined that P. bedriagae secretion exhibits antimicrobial properties, and many proteins with different molecular masses were identified in the skin secretion.</p><p><strong>Discussion: </strong>There are no skin secretion studies on <i>P. bedriage</i> in the literature. However, activity studies were also conducted on the skin secretion of another Pelophylax species, P. ridibundus. The study examined the antibacterial activities of the skin secretion against some gram-positive and gram-negative bacteria using a plate well diffusion assay. Their study showed that E. coli formed a 21 mm zone and S. aureus formed a 24 mm zone.</p><p><strong>Conclusion: </strong>This study is preliminary, and in future studies, AMPs in skin secretions can be isolated by chromatographic methods, such as HPLC, and peptides can be sequenced and identified in detail.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"650-656"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145233265","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}

{"title":"Optimizing Sleep in Athletes: The Potential of α-Lactalbumin in Nutrition Intervention.","authors":"Jingjing Li, Xuepeng Bian","doi":"10.2174/0109298665363873250623103811","DOIUrl":"10.2174/0109298665363873250623103811","url":null,"abstract":"<p><p>Athletes frequently encounter sleep deprivation due to the demands of high-intensity training and competition, which can significantly impair their physical recovery and athletic performance. α-Lactalbumin (α-LA), a key component of whey protein that is rich in tryptophan, has been shown to promote the synthesis of serotonin and melatonin, thereby regulating sleep cycles. Moreover, α-LA has demonstrated the ability to reduce inflammation and oxidative stress associated with fatigue and stress, further contributing to improved sleep quality. This review provides a critical evaluation of the current evidence supporting the role of α-LA in enhancing sleep quality in athletes through mechanisms such as neurotransmitter regulation, immune function improvement, and enhancement of antioxidant defenses. Additionally, it highlights the necessity for further research into the differential effects of α -LA on sleep across various sports and gender groups, as well as its potential synergistic interactions with other nutrients. These insights are essential for developing optimized nutritional interventions aimed at enhancing athletic performance.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"402-413"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12678987/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144637802","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}

{"title":"Inhibition of Canonical Wnt Signaling in Renal Cell Carcinoma Bone Metastasis: An Immunohistochemical Analysis of DKK1 and LRP5 Expression.","authors":"Zixiong Huang, Yiqing Du, Huaqi Yin, Gongwei Wang, Tao Xu","doi":"10.2174/0109298665357331250416081850","DOIUrl":"10.2174/0109298665357331250416081850","url":null,"abstract":"<p><strong>Background and objective: </strong>Canonical Wnt (Wnt/β-catenin) signaling maintains bone homeostasis by promoting osteoblastic activities. The inhibitory factor, Dickkopf-1 (DKK1), enhances bone resorption in malignant diseases. Low-density lipoprotein-related protein (LRP) 5 is antagonized by DKK1. This study aimed to investigate the expression of DKK1 and LRP5 in renal cell carcinoma bone metastasis (RCC-BM).</p><p><strong>Methods: </strong>RCC-BM patients with paired samples of primary and metastatic lesions were selected for the study (RCC-BM group). RCC patients without any metastasis served as the control group (RCC-only group). Immunohistochemical staining with monoclonal anti-DKK1 and polyclonal anti- LRP5 antibody was conducted on paraffin-embedded slides. The staining results were recorded using scoring according to staining intensity in the renal tissue adjacent to the tumor, primary RCC lesions, and RCC-BM lesions.</p><p><strong>Results: </strong>DKK1 was differently expressed among normal renal tissues, primary RCC, and RCC- BM tissues (p<0.001). The DKK1 expression in primary RCC was significantly lower than that in normal renal tissues (p<0.001) without a difference between the RCC-BM and RCC-only groups. DKK1 expression in bone metastasis was significantly higher than that in primary tumors (p<0.001). For RCC-BM patients, the expression of LRP5 in the primary tumor was significantly lower than that in adjacent renal tissues (p<0.01). The tendency of lower expression was found in primary RCC from RCC-BM patients compared to RCC without metastasis (p=0.073).</p><p><strong>Conclusion: </strong>A \"rebound\" pattern of DKK1 expression in bone metastasis lesions and the decreasing LRP5 expression in primary lesions of RCC-BM patients suggested that Wnt/β-catenin signaling was inhibited in RCC-BM. The overexpression of DKK1 and reduced expression of LRP5 suggest that these markers may be useful for the early prediction of RCC-BM.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"327-334"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144028841","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}

{"title":"Molecular Cloning and Expression of Cryptocyanin Gene Isolated from an Indian Variety of <i>Scylla olivacea</i>.","authors":"Simran Mann, Chittibabu Shanthi, Manu Asthana","doi":"10.2174/0109298665387863250506105601","DOIUrl":"10.2174/0109298665387863250506105601","url":null,"abstract":"<p><strong>Background: </strong>Molting and reproduction play vital roles in the life cycle of brachyuran crabs, and these two processes are closely interconnected. A key player in the molting cycle is cryptocyanin, which is similar to hemocyanin in sequence, size, and structure. Hemocyanin is a copper-containing oxygen-binding protein, while cryptocyanin is a copper-free protein that lacks oxygen-binding capacity.</p><p><strong>Objective: </strong>The goal of the study was to carry out the isolation, cloning, and expression of the partial cryptocyanin gene from the Indian variety of <i>Scylla olivacea</i>.</p><p><strong>Methods: </strong>The partial cryptocyanin gene was isolated from the hemocytes of the <i>S. olivacea</i> male and female crabs by qPCR for comparative expression analysis of the cryptocyanin gene.</p><p><strong>Results: </strong>We successfully amplified, cloned, and expressed a 519 bp partial cDNA encoding cryptocyanin from the Indian variety of<i> Scylla olivacea</i>, within the pRSET-B vector.</p><p><strong>Discussion: </strong>In this study, we conducted a comprehensive analysis of cryptocyanin expression in male and female crabs during the intermolt stage. Our findings revealed Cq values of 28.97 for males and 33.68 for females, highlighting a significantly lower abundance of cryptocyanin protein in female crabs.</p><p><strong>Conclusion: </strong>Our study showed that crustacean cDNA can be effectively expressed in bacterial vectors, and clones were stable for up to 6 months at -80oC. Real-time data showed a significant difference in cryptocyanin levels between male and female crabs. This finding highlights the need for further research with a larger sample size for better understanding.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"368-375"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144143106","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}

{"title":"Unveiling the Potential Role of Hesperetin and Emodin as a Combination Therapy to Inhibit the Pancreatic Cancer Progression against the C-Met Gene.","authors":"Rangaraj Kaviyaprabha, Thandaserry Vasudevan Miji, Puthupparambil Shaji Sreelakshmi, Sridhar Muthusami, Palanisamy Arulselvan, Muruganantham Bharathi","doi":"10.2174/0109298665363165250225100109","DOIUrl":"10.2174/0109298665363165250225100109","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Pancreatic adenocarcinoma (PAAD) is one of the most prevalent cancers, and it has high death rates. Only 10% of PAAD patients can survive until 5 years. Hence, the improvement of survival rate of the patients should be improved.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Aim: &lt;/strong&gt;The present study used a computational approach to identify novel biomarkers and potentially effective small drug-like molecules in PAAD.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Objective: &lt;/strong&gt;The objective of this study was to identify the Differentially Expressed Genes (DEGs) and survival rate affecting genes (SDEGs) to single out the specific gene responsible for pancreatic cancer and predict the efficacy of interactions with hesperetin and emodin. Further, another objective was to validate the predicted efficacies using an MTT assay.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;The GEPIA2 database was used to analyze the TCGA-PAAD dataset and identify DEGs and SDEGs. Venn identified the commonly scattered genes between the DEGs and SDEGs. Network Analyst v3.0, CytoScape v3.10.1, and cytoHubbawere used to construct protein-protein interactions (PPI) network and identifying hub genes which were described as target proteins. The Protein Data Bank (PDB) and PubChem were utilized to obtain the PDB structure of the target proteins and 13 phytocompounds in SDF format. Molecular docking studies were carried out and visualized by utilizing Autodock vina and Discovery Studio Visualizer v19.1.0.1828. The cytotoxicity was measured in the MiaPaCa-2 cell line after being treated with hesperetin and emodin.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;A total of 9219 Differentially Expressed Genes (DEGs) from the TCGA-PAAD dataset were identified. Among them, 8740 and 479 genes were up and down-regulated with the statistical significance of P ≤ 0.05, respectively. Likely, 500 most survival rate affecting genes (SDEGs) in PAAD patients with a statistical significance of P ≤ 0.05 were identified. The common 137 genes were identified between these obtained DEGs and SDEGs. The survival heat map was delineated for the predicted 137 common genes. Ninety-six genes were identified as the most hazardous genes (highlighted in red). After that, the network was constructed by using PPI for the most hazardous 96 genes. From the constructed PPI network, the highly interacted top 10 genes were identified. The survival analysis was carried out to identify the most hazardous genes and revealed that all the identified genes significantly reduced the survival rate of the patients affected by PAAD. From that, high survival affecting 5 genes, such as CDK1, CENPE, NCAPG, KIF20A, and c-MET, were selected for further analysis. The molecular docking studies were carried out for the identified top 5 genes, with the 13 phytocompounds reviewed previously for anti-- cancer activity. The molecular docking analysis revealed that the hesperetin (binding affinity (BA) = -8.0 kcal/mol; Root mean square deviation (RMSD) = 2.012 Å) and emodin (BA = -8.6 k","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"280-298"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143701385","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}

{"title":"miR-584-5p Regulates MSMO1 to Modulate the AKT/PI3K Pathway and Inhibit Breast Cancer Progression.","authors":"Xin Li, Jie Liu, Lili He, Mi Tian, Yingying Xu, Bing Peng","doi":"10.2174/0109298665339026250114070523","DOIUrl":"10.2174/0109298665339026250114070523","url":null,"abstract":"<p><strong>Introduction: </strong>Endogenous microRNAs (miRNAs) are critical regulators of tumor progression, making their role in breast cancer an important area of investigation.</p><p><strong>Methods: </strong>This study examined the regulation of MSMO1 by miR-584-5p in breast cancer cells. Using bioinformatics and Western blotting, we confirmed MSMO1 expression in breast cancer cells and evaluated its effects on cell migration, invasion, and the AKT signaling pathway. In vivo experiments further supported these findings. The interaction between miR-584-5p and MSMO1 was validated through luciferase reporter assays, while functional studies highlighted the impact of miR-584-5p on cancer progression.</p><p><strong>Results: </strong>Our findings revealed that MSMO1 is upregulated in breast cancer, enhancing cell migration and invasion. Silencing MSMO1 diminished AKT pathway activity, and luciferase assays confirmed MSMO1 as a direct target of miR-584-5p.</p><p><strong>Conclusion: </strong>Overexpression of miR-584-5p suppressed migration and invasion of breast cancer cells. In summary, miR-584-5p is likely to modulate MSMO1 and subsequently regulate the AKT/ PI3K pathway, presenting a promising therapeutic target for breast cancer treatment.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"171-182"},"PeriodicalIF":1.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143415087","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}

{"title":"Enhancing Tissue Factor Production: The Role of N-Glycosylation and ERAD Pathway Modulation.","authors":"Yi-Shi Liu, Yue Dou, Xiaoman Zhou, Zijie Li, Nakanishi Hideki","doi":"10.2174/0109298665364078250519065417","DOIUrl":"10.2174/0109298665364078250519065417","url":null,"abstract":"<p><strong>Background: </strong>Tissue Factor (TF) is a crucial transmembrane glycoprotein that triggers blood coagulation upon vascular or tissue injury by binding to plasma factors VII and VIIa. In recent years, the demand for TF has rapidly increased due to its pivotal role in preoperative coagulation tests. However, large-scale production of TF remains challenging despite successful recombinant expression, as incorrect post-translational modifications adversely affect TF activity.</p><p><strong>Objective: </strong>This study aims to investigate the role of post-translational modifications, specifically N-glycosylation, in TF activity and stability. Additionally, it explores strategies to enhance TF production by reducing its degradation through genetic modulation.</p><p><strong>Methods: </strong>We compared TF activity derived from human cells and E. coli to assess the impact of post-translational modifications. Furthermore, we examined the effect of N-glycosylation on TF function. To address TF degradation, we knocked out the HRD1 gene, a key component of the endoplasmic- reticulum-associated degradation (ERAD) pathway, and evaluated its impact on TF stability and activity.</p><p><strong>Results: </strong>TF produced in human cells exhibited higher activity than TF expressed in E. coli, emphasizing the importance of post-translational modifications. Specifically, N-glycosylation was found to influence TF activity and stability. Additionally, we observed that knocking out the HRD1 gene effectively reduced TF degradation without compromising its activity.</p><p><strong>Conclusion: </strong>Our findings underscore the crucial role of N-glycosylation in TF function and stability. Moreover, the modulation of the ERAD pathway through knocking out HRD1 presents a promising approach for enhancing TF production. These insights could contribute to the large-scale manufacturing of functionally active TF for clinical and research applications.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"508-518"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187769","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}

{"title":"Purification, Crystallization, and Preliminary X-ray Diffraction Studies on Hemoglobin from the Angora Goat (<i>Capra Aegagrus Hircus)</i>.","authors":"Farheen Pervez, Shobana Nagaraj, Mpho Setshedi, Arshad Mather, Ponnuswamy Mondikalipudur Nanjappa Gounder, Yasien Sayed, Ramesh Pandian","doi":"10.2174/0109298665370730250708065019","DOIUrl":"10.2174/0109298665370730250708065019","url":null,"abstract":"<p><strong>Introduction: </strong>Angora goats are a distinct breed that differs significantly from common goats and shares a similar appearance to sheep. In Angora goats, only the level of glutathione (GSH) is elevated during under-stimulated conditions, as well as after the period of hypoxic stress; however, no changes are found in 2,3-diphosphoglycerate (2,3-DPG) levels, which are commonly present in the red blood cells (RBCs) of most mammals. We chose the Angora goat for our investigation because no previous studies have been conducted on the structural and functional aspects of hemoglobin (Hb). In addition, no sequence or structural information is currently available in any database.</p><p><strong>Methods: </strong>Angora goat Hb was isolated and purified by anion-exchange chromatography, followed by crystallization using various methods. X-ray data collection for Angora goat Hb was performed under a liquid nitrogen cryo-stream using a <i>Bruker D8 Venture Bio Photon III 28-pixel array area detector</i> system.</p><p><strong>Results: </strong>Good diffracting crystals were obtained using the hanging-drop vapor-diffusion method with polyethylene glycol (PEG) 3350 as the precipitant in water, without the addition of any salt or buffer. The Angora goat Hb diffracted to a resolution of 1.85 Å, and the structure solution was obtained by the molecular replacement method, using the structure of domestic goat Hb as the starting model.</p><p><strong>Discussion: </strong>The solved structure of Angora goat crystallized in the monoclinic space group P21, consisting of one whole biological molecule in the asymmetric unit, with unit cell dimensions of a = 52.08 Å, b = 76.70 Å, c = 74.08 Å, and β = 91.77 °. The solvent content and Matthews coefficient (Vm) for the Angora goat Hb are 49.05% and 2.41 ų/Da, respectively, and are within the normal range for protein crystals.</p><p><strong>Conclusion: </strong>Purification, crystallization, and preliminary X-ray diffraction studies of Angora goat Hb were performed successfully. Structural refinement and biophysical characterization of Angora goat Hb are in progress in the absence and presence of GSH and 2,3-DPG.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"519-524"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144637804","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}

{"title":"Improving Acid-Base Pair Concentration in Wash/Elution Buffer Eliminates Elution Peak-Shouldering in Cation Exchange Chromatography.","authors":"Fei Huang, Na Liu","doi":"10.2174/0109298665394172250926073117","DOIUrl":"10.2174/0109298665394172250926073117","url":null,"abstract":"<p><strong>Introduction: </strong>Peak-shouldering elution behavior was a common and unexpected result in bind-and-elute mode Cation Exchange Chromatography (CEX), which may be due to the pH transition during the elution step and the aggregation tendency of target proteins.</p><p><strong>Methods: </strong>Improving the concentration of acid-base pairs in the wash buffers or elution buffers without changing pH or conductivity effectively resolved the peak-shouldering issue in CEX.</p><p><strong>Results: </strong>In the case of molecule A, the shoulder peak was eliminated in the CEX run by increasing the NaAc-HAc concentration from 50 mM to 100 mM in the elution buffer or from 50 mM to 75 mM in the wash buffer. Higher NaAc-HAc concentrations affect the pH transition in the early stages of the elution step, which may explain the elimination of the shoulder peak. A similar result was observed for molecule B, where increasing the Tris-HCl concentration in the elution buffer from 50 mM to 80 mM also removed the shoulder peak during elution.</p><p><strong>Discussion: </strong>The successful elimination of peak-shouldering behavior by increasing acid-base pair concentrations highlights the critical role of buffer capacity in modulating pH transitions during CEX. While this strategy offers a simple and effective solution, further investigation is needed to assess its applicability across diverse protein types and buffer systems.</p><p><strong>Conclusion: </strong>These results demonstrate that increasing the concentration of acid-base pairs in the elution buffer or wash buffer of CEX using NaAc-HAc or Tris-HCl buffers is an effective strategy for eliminating the shoulder-peak.</p>","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":"711-717"},"PeriodicalIF":1.1,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145233300","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}