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Capturing G protein-coupled receptors into native lipid-bilayer nanodiscs using new diisobutylene/maleic acid (DIBMA) copolymers 利用新型二异丁烯/马来酸（DIBMA）共聚物将G蛋白偶联受体捕获到天然脂质双层纳米圆盘中。
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-09-04 DOI: 10.1016/j.ymeth.2025.08.013
Ci Chu , Carolyn Vargas , Maria Carolina Barbosa , Simon Sommerhage , Gerald N. Rechberger , David Pahovnik , Ema Žagar , Gunnar F. Schröder , Sandro Keller , Manuel Etzkorn
Many membrane proteins, including G protein-coupled receptors (GPCRs), are susceptible to denaturation when extracted from their native membrane by detergents. Therefore, alternative methods have been developed, including amphiphilic copolymers that enable the direct extraction of functional membrane proteins along with their surrounding lipids. Among these amphiphilic copolymers, styrene/maleic acid (SMA) and diisobutylene/maleic acid (DIBMA) polymers have been extensively studied. Despite their many benefits, SMA and DIBMA polymers also have considerable drawbacks limiting their applications. Herein, we describe a series of new amphiphilic copolymers derived from DIBMA via partial amidation of the carboxylate pendant groups with various biocompatible amines. We characterize the new polymer’s nanodisc-forming properties and ability to extract the melanocortin 4 receptor (MC4R), a prototypical class A GPCR. While each new DIBMA variant displays features that may be favorable for selected applications, we identified a PEGylated DIBMA variant called mPEG4-DIBMA as particularly promising. In the tested system mPEG4-DIBMA abolishes unspecific interactions and outperforms other polymers by achieving higher extraction efficiencies of MC4R from Sf9 insect cell membranes. The new nanodisc-forming polymer combines two key advantages that are crucial for investigating GPCRs in a well-defined but still native lipid-bilayer environment, thus paving the way for manifold future applications.
许多膜蛋白，包括G蛋白偶联受体（gpcr），当被洗涤剂从其天然膜中提取时，容易变性。因此，替代方法已经开发出来，包括两亲性共聚物，可以直接提取功能膜蛋白及其周围的脂质。在这些两亲共聚物中，苯乙烯/马来酸（SMA）和二异丁烯/马来酸（DIBMA）聚合物得到了广泛的研究。尽管SMA和DIBMA聚合物有许多优点，但它们也有相当大的缺点，限制了它们的应用。在此，我们描述了一系列新的两亲性共聚物，这些共聚物是通过羧酸悬垂基团与各种生物相容性胺的部分酰胺化而得到的。我们表征了新聚合物的纳米圆盘形成特性和提取黑素皮质素4受体（MC4R）的能力，这是一种典型的a类GPCR。虽然每个新的DIBMA变体都显示出可能有利于选定应用的特征，但我们确定了一个称为mPEG4-DIBMA的聚乙二醇化DIBMA变体特别有前途。在测试的体系中，mPEG4-DIBMA消除了非特异性相互作用，并通过从Sf9昆虫细胞膜中获得更高的MC4R提取效率而优于其他聚合物。这种新型纳米盘状聚合物结合了两个关键优势，这对于在定义明确但仍然是天然脂质双分子层环境中研究gpcr至关重要，从而为未来的多种应用铺平了道路。
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引用次数: 0
Efficient RNA nucleotide encoding enhances the accurate prediction of ac4C modifications 高效的RNA核苷酸编码增强了对ac4C修饰的准确预测。
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-31 DOI: 10.1016/j.ymeth.2025.07.012
Na Li , Xiao Wang , Ming Zeng , Feng Cao , Ke Qiu , Jianbo Qiao
RNA N4-acetylcytidine (ac4C) modification plays a vital role in gene regulation and cellular function. Accurate identification of ac4C sites is essential for elucidating their biological significance. However, existing prediction methods struggle to capture complex sequence patterns, limiting their accuracy. To address this, we propose GO-ac4C, an efficient prediction framework that integrates byte-pair encoding with nucleotide compositional features. GO-ac4C employs dynamic byte-pair encoding to learn optimal subsequence representations and enhances them with compositional features to effectively capture key motifs in RNA sequences. Experimental results demonstrate that GO-ac4C significantly outperforms state-of-the-art methods across multiple evaluation metrics and offers new insights into the mechanisms of RNA modification.
RNA n4 -乙酰胞苷（ac4C）修饰在基因调控和细胞功能中起着至关重要的作用。准确鉴定ac4C位点对于阐明其生物学意义至关重要。然而，现有的预测方法难以捕捉复杂的序列模式，限制了它们的准确性。为了解决这个问题，我们提出了GO-ac4C，一个有效的预测框架，集成了字节对编码和核苷酸组成特征。GO-ac4C采用动态字节对编码来学习最优子序列表示，并用组合特征对其进行增强，从而有效捕获RNA序列中的关键基序。实验结果表明，GO-ac4C在多个评估指标上明显优于最先进的方法，并为RNA修饰机制提供了新的见解。
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引用次数: 0
Rapid in vitro and computer-aided method for assessing synergistic interactions between NSAIDs and analgesics 快速体外和计算机辅助方法评估非甾体抗炎药和镇痛药之间的协同相互作用。
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-30 DOI: 10.1016/j.ymeth.2025.08.012
Fernando Silva , Gustavo Costa , Francisco Veiga , Vera Moura , Francisca Dias , Fátima Cerqueira , Rui Medeiros , Ana Cláudia Paiva-Santos
Pain is a complex phenomenon that plays a significant role in various diseases, influencing both the physical and psychological well-being of individuals. In clinical practice, combining nonsteroidal anti-inflammatory drugs (NSAIDs) with analgesics, such as paracetamol or metamizole, has become a widely adopted strategy to manage pain. Although the synergistic effects of combining NSAIDs with analgesics are well recognized in clinical practice, this approach is primarily based on empirical clinical experience. Our work aims to present a rapid method for evaluating the anti-inflammatory effects of drug combinations through in vitro assays combined with computer-aided data processing and analysis.
We conducted two simple and rapid in vitro assays, the Griess and DPPH assays, to evaluate the effects of NSAID–analgesic combinations and demonstrate their synergistic interactions, using the free web application SynergyFinder Plus. This computer-aided analysis enabled a quantitative assessment of drug interactions, enhancing the interpretation of the experimental data. Furthermore, to better understand the results obtained from previous experiments, we analysed the anti-inflammatory effects of ketoprofen and dexketoprofen in combination with metamizole and paracetamol through quantitative real-time PCR (qRT-PCR). Our findings reveal synergistic interactions between NSAIDs and analgesics in terms of their anti-inflammatory and antioxidant activities.
This work could be the first step for the study of the mechanisms behind the synergistic interactions between NSAIDs and analgesics for the treatment of pain, mainly when inflammatory processes are involved. Consequently, this study aims to contribute to the exploration of non-opioid drug combinations, addressing the urgent need for alternative analgesic strategies that minimize opioid use.
疼痛是一种复杂的现象，在各种疾病中起着重要作用，影响着个体的生理和心理健康。在临床实践中，非甾体抗炎药（NSAIDs）与镇痛药（如扑热息痛或metamizole）联合使用已成为一种广泛采用的治疗疼痛的策略。尽管非甾体抗炎药与镇痛药联合使用的协同作用在临床实践中得到了广泛认可，但这种方法主要基于临床经验。我们的工作旨在通过体外试验结合计算机辅助数据处理和分析，提出一种快速评估药物联合抗炎作用的方法。我们使用免费的网络应用程序SynergyFinder Plus进行了两种简单快速的体外试验，Griess和DPPH试验，以评估nsaid -镇痛药联合使用的效果，并证明它们之间的协同作用。这种计算机辅助分析能够对药物相互作用进行定量评估，增强对实验数据的解释。此外，为了更好地理解之前的实验结果，我们通过实时荧光定量PCR （qRT-PCR）分析了酮洛芬和右酮洛芬与metamizole和paracetamol联合使用的抗炎作用。我们的发现揭示了非甾体抗炎药和镇痛药在抗炎和抗氧化活性方面的协同相互作用。这项工作可能是研究非甾体抗炎药和镇痛药治疗疼痛的协同相互作用机制的第一步，主要是在炎症过程中。因此，本研究旨在促进非阿片类药物组合的探索，解决替代镇痛策略的迫切需要，最大限度地减少阿片类药物的使用。
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引用次数: 0
Validation of combo ichroma as a reliable concentration-based alternative for AST and ALT measurement in liver disease monitoring 联合色度作为肝脏疾病监测中AST和ALT测量的可靠的基于浓度的替代方法的验证
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-27 DOI: 10.1016/j.ymeth.2025.08.011
Minsoo Kim , Su A Kim , Jeong Min Kim , Hee Young Kim , Ho Yeong Yoon , Sung Won Park , Daegyun Park , Ji Sook Han , Ki Tae Suk

Background

Traditional assays for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) measure enzymatic activity, which degrades during long-term frozen storage, threatening the accuracy of the assay. Instead, Combo ichroma (CI), a fluorescence immunoassay that quantifies AST and ALT concentrations, is a robust alternative for retrospective and point-of-care liver function testing, free from the influence of long-term storage.

Methods

Serum samples from 256 individuals (controls and patients with hepatitis, cirrhosis, or liver cancer) were collected and stored at −80 °C for an average of three years. AST and ALT were measured using CI, the 7180 clinical analyzer (HT), and Atellica CH 930 analyzer performed immediately after collection (HL). Correlations between AST, ALT, and AST/ALT ratios were analyzed. Random Forest Regression (RFR) models using CI or HT data were developed to predict HL-derived AST/ALT ratios.

Results

CI-AST showed strong correlation with HL-AST across all groups (R2 > 0.95), outperforming HT-AST, especially in controls (R2 = 0.58). CI-ALT moderately correlated with HL-ALT (R2 = 0.87), surpassing HT-ALT (R2 = 0.71). AST/ALT ratios varied across methods due to ALT variability, but RFR using CI data accurately predicted HL ratios (R2 = 0.85–0.91). Subgroup analysis confirmed CI’s superior concordance across etiologies.

Conclusions

CI enables activity-independent, reliable measurement of AST and ALT even after extended storage, outperforming enzymatic assays in precision and correlation. Its simplicity, and compatibility with machine learning models position CI as a promising tool for liver enzyme diagnostics in both clinical and resource-limited settings.
传统的天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）检测方法测量的是酶活性，在长期冷冻储存过程中酶活性会降低，从而影响检测的准确性。相反，组合染色法（CI）是一种定量AST和ALT浓度的荧光免疫分析法，是回顾性和即时肝功能检测的可靠替代方法，不受长期储存的影响。方法收集256例（对照组和肝炎、肝硬化或肝癌患者）的血清样本，在- 80℃平均保存3年。采用CI、7180临床分析仪（HT）和Atellica CH 930采集后立即测定AST和ALT。分析AST、ALT及AST/ALT比值的相关性。随机森林回归（RFR）模型使用CI或HT数据来预测hl衍生的AST/ALT比率。结果sci - ast与HL-AST在各组间均表现出较强的相关性（R2 > 0.95），优于HT-AST （R2 = 0.58）。CI-ALT与HL-ALT中度相关（R2 = 0.87），优于HT-ALT （R2 = 0.71）。由于ALT的可变性，不同方法的AST/ALT比率不同，但使用CI数据的RFR准确预测HL比率（R2 = 0.85-0.91）。亚组分析证实了CI在病因上的优越一致性。结论sci能够独立于活性，可靠地测量AST和ALT，即使在延长储存后，在精度和相关性方面优于酶分析。它的简单性和与机器学习模型的兼容性使CI在临床和资源有限的情况下都成为肝酶诊断的有前途的工具。
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引用次数: 0
Optimized CUT&Tag enables robust epigenome profiling in Schizosaccharomyces pombe 优化的CUT&Tag能够在裂糖菌pombe中实现稳健的表观基因组分析
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-22 DOI: 10.1016/j.ymeth.2025.08.010
Cheng-Zhi Huang , Kang-Di Zhou , Wei Ma
We optimized permeabilization for CUT&Tag in S. pombe, enabling robust H3K9me3 profiling using lightly fixed permeabilized sepheroplasts, overcoming limitations of ChIP-seq including crosslinking artifacts and high cell input. We established an optimized Cleavage Under Targets and Tagmentation (CUT&Tag) protocol for high-resolution epigenome profiling in Schizosaccharomyces pombe using Critical permeabilization refinements identified Lywallzyme as the optimal enzyme for spheroplast generation (>95 % efficiency in 60 min at 10 mg/mL), outperforming Zymolyase-20 T and combinatorial treatments. Systematic parameter optimization revealed concentration-dependent digestion kinetics and an inverse cell load-efficiency relationship (5 × 105 cells achieving > 90 % conversion in 50 min at 5 mg/mL). Validated through H3K9me3 mapping in wild-type and clr4Δ strains (10⁶ cells/replicate), this approach captured specific heterochromatic enrichment at centromeres/telomeres with complete signal ablation in mutants, while reduced spike-in DNA (0.2 pg) significantly enhanced signal-to-noise ratios. The protocol enables robust epigenomic analysis with minimal cell input and enhanced resolution.
我们优化了S. pombe中CUT&；Tag的渗透性，使用轻度固定的渗透性磷脂质体实现稳健的H3K9me3分析，克服了ChIP-seq的局限性，包括交联伪影和高细胞输入。我们建立了一个优化的切割靶下和标记（CUT&Tag）方案，用于高分辨率的裂糖酵母表观基因组分析，使用临界渗透细化鉴定出Lywallzyme是球质体生成的最佳酶（在10 mg/mL条件下60分钟效率为95%），优于酶解酶- 20t和组合处理。系统参数优化揭示了浓度依赖的消化动力学和反向的细胞负载效率关系（5 × 105个细胞在5 mg/mL的条件下在50分钟内达到90%的转化率）。通过在野生型和clr4Δ菌株（10 26个细胞/重复）中进行H3K9me3定位验证，该方法在突变体中捕获了着丝粒/端粒的特异性异染色质富集，信号完全消失，同时减少了DNA尖峰（0.2 pg），显著提高了信噪比。该方案能够以最小的细胞输入和增强的分辨率进行稳健的表观基因组分析。
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引用次数: 0
Volumetric dried blood spot microsampling: A sustainable, patient-friendly, and practical approach for retinol and α-tocopherol analysis in a clinical setting 体积干燥血斑微采样：一个可持续的，病人友好的，实用的方法视黄醇和α-生育酚分析在临床设置
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-22 DOI: 10.1016/j.ymeth.2025.08.006
Chaweewan Suwanvecho , Lenka Kujovská Krčmová , František Švec
Dried blood spot (DBS) technique has gained significant attention due to the growth of decentralized diagnostics. This technique reduces the number of hospital visits for patients and the workload for personnel in specialized hospitals. This microsampling method provides an environmentally friendly (green) and patient-friendly alternative to conventional phlebotomy. Challenges related to sample heterogeneity in traditional DBS cards have been overcome by the volumetric DBS sampling using new types of commercially available devices. Due to the unstable nature of the analytes, commercial volumetric DBS devices allow blood sampling at primary care units in remote settings and facilitate transport it via temperature-controlled systems. Blood sample stability has improved from 24 h at 4–8 °C to 30 days at −80°C. DBS also requires over 1000 times less shipping and storage space than liquid blood. We optimized the DBS method to require only 10 µL of blood and achieve extraction efficiencies of over 90 % for retinol when the result from validated method is the reference value. However, α-tocopherol recovery varied from 53 to 75 % depending on the filter paper type used. Furthermore, we successfully developed a liquid–liquid extraction method for both analytes from whole blood, with over 90 % recovery. Our approaches eliminate the need for separate serum and erythrocyte extractions, simplify sample preparation, and reduce reagent use and energy consumption. Both devices enable reliable volumetric collection. Our approach makes micronutrient monitoring more accessible and enables sample collection in decentralized settings. This aligns with the objectives of green analytical chemistry and universal health coverage.
由于分散式诊断的发展，干血斑（DBS）技术得到了极大的关注。这种技术减少了患者的医院就诊次数和专科医院工作人员的工作量。这种微采样方法提供了一种环境友好（绿色）和病人友好的替代传统的静脉切开术。传统DBS卡中与样本异质性相关的挑战已经被使用新型商用设备的容量DBS采样所克服。由于分析物的不稳定性，商用容量DBS设备允许在偏远地区的初级保健单位进行血液采样，并便于通过温控系统运输。血液样品的稳定性从4-8°C下的24小时提高到- 80°C下的30天。DBS所需的运输和储存空间也比液体血液少1000多倍。我们优化了DBS方法，只需要10µL的血液，当验证方法的结果为参考值时，视黄醇的提取效率超过90%。然而，α-生育酚的回收率根据滤纸类型的不同，从53%到75%不等。此外，我们成功地开发了一种液-液萃取方法，可从全血中提取这两种分析物，回收率超过90%。我们的方法消除了分离血清和红细胞提取的需要，简化了样品制备，减少了试剂的使用和能源消耗。这两种设备都支持可靠的体积收集。我们的方法使微量营养素监测更容易获得，并能够在分散的环境中收集样本。这符合绿色分析化学和全民健康覆盖的目标。
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引用次数: 0
Coffee – a ubiquitous substitute for uranyl acetate in staining of biological ultrathin sections for electron microscopy studies 咖啡-在电子显微镜研究的生物超薄切片染色中，醋酸铀酰的普遍替代品
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-21 DOI: 10.1016/j.ymeth.2025.08.009
Claudia Mayrhofer , Robert Zandonella , Willi Salvenmoser , Ilse Letofsky-Papst
The use of uranyl acetate, a staining agent successfully used for decades in electron microscopy of biological specimens, is now strictly regulated by law due to its toxicity and radioactivity. It is even banned in some laboratories. In the meantime, there are a number of substitutes on the market, none of which comes close to the very good staining results of uranyl acetate, or only partially, and some of which are also toxic. In this paper, two alternatives to uranyl acetate are presented, namely coffee, which is used in countless households, and pure chlorogenic acid, which is a component of coffee. We used the well-known zebrafish as biological test material, focusing on the mitochondrial membranes. The staining ability of coffee and chlorogenic acid compared with commercially available staining agents as well as uranyl acetate is assessed by the interference contrast between membranes and their environment. This work also describes how a subjective impression of good or bad contrast can be cast into an objective and comparable numerical value.
醋酸铀酰是一种在生物标本电子显微镜中成功使用了几十年的染色剂，由于其毒性和放射性，现在受到法律的严格管制。一些实验室甚至禁止使用。与此同时，市场上有许多替代品，没有一种接近醋酸铀酰的非常好的染色效果，或者只是部分，其中一些也是有毒的。本文介绍了醋酸铀酰的两种替代品，即在无数家庭中使用的咖啡和纯绿原酸，这是咖啡的一种成分。我们用众所周知的斑马鱼作为生物测试材料，重点研究线粒体膜。通过膜与环境的干涉对比，比较了咖啡和绿原酸与市售染色剂以及醋酸铀酰的染色能力。这项工作还描述了一个主观印象的好或坏的对比可以投射到一个客观的和可比较的数值。
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引用次数: 0
Exploiting Anisotropic Orientation in Nanoparticle-Aided Cryo-Electron Microscopy Sampling 利用纳米粒子辅助低温电子显微镜取样的各向异性取向
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-21 DOI: 10.1016/j.ymeth.2025.08.008
Yeeun Kim , Changin Kim , Yongsoo Yang , Hyotcherl Ihee
Measuring the conformational distribution of small proteins is essential to understanding their role in biological systems. Multi-Tilt Nanoparticle-aided cryo-electron microscopy sampling (MT-NACS) was devised to measure the three-dimensional interparticle distance distribution (P(d)) of two gold nanoparticles (AuNPs) labeled on a protein by taking cryogenic electron microscopy (cryo-EM) images at multiple-tilt angles. However, tracking the same particles in a pseudo-tomographic manner during the multi-tilt cryo-EM experiments and data analysis requires extensive time and effort. Here, we report that proper incorporation of AuNP pair angle distribution allows reliable determination of the P(d) only using the cryo-EM images collected at a single tilt angle. The trends of structural changes in calmodulin (CaM) measured by single-tilt angle NACS (ST-NACS) and MT-NACS are consistent, as the tendencies of changes in the P(d) of AuNP-labeled CaM measured by both methods are similar. Our approach provides an efficient tool for measuring the conformational distribution and structural transition of small proteins.
测量小分子蛋白质的构象分布对于理解它们在生物系统中的作用至关重要。设计了多倾斜纳米粒子辅助低温电子显微镜采样（MT-NACS），通过在多个倾斜角度拍摄低温电子显微镜（cro - em）图像来测量标记在蛋白质上的两个金纳米粒子（AuNPs）的三维粒子间距离分布（P(d)）。然而，在多倾斜低温电镜实验和数据分析中，以伪层析方式跟踪相同的粒子需要大量的时间和精力。在这里，我们报告了适当地结合AuNP对角度分布可以可靠地确定P(d)，仅使用在单一倾斜角度下收集的冷冻电镜图像。单倾角NACS （ST-NACS）和MT-NACS测量的钙调素（CaM）结构变化趋势是一致的，因为两种方法测量的aunp标记CaM的P(d)变化趋势相似。我们的方法为测量小分子蛋白质的构象分布和结构转变提供了一种有效的工具。
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引用次数: 0
PCR-generated DNA templates enable efficient, rapid, and cost-effective mRNA synthesis pcr生成的DNA模板能够高效、快速、经济地合成mRNA。
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-16 DOI: 10.1016/j.ymeth.2025.08.007
Naveen Kumar , Nicholas C. Hazell , Jiani Bei , Tina Nguyen , Haitao Hu
In vitro transcription (IVT) is a widely used technique for mRNA synthesis in both basic research and the development mRNA-based vaccines and therapies. The efficiency of IVT critically depends on the quality and integrity of the linear DNA templates. The conventional method for template DNA preparation involves plasmid propagation in bacteria followed by enzymatic linearization, which is labor-intensive and costly. Here, we describe a cell-free, PCR-based approach for generating high-quality, high-yield linear DNA templates. We extensively compared the PCR-based method with the conventional plasmid-based approach in terms of IVT efficiency, mRNA production, and the immunogenicity of the resulting mRNA-LNP (lipid nanoparticle) vaccines. Compared to the plasmid-derived DNA, the PCR-based method yielded higher amounts of both DNA templates and transcribed mRNA, while maintaining mRNA quality and integrity. Importantly, mRNA-LNP vaccines encoding the SARS-CoV-2 spike protein, generated from both methods, elicited robust and comparable immune responses in mice, with no significant differences observed between the two template methods. Our findings highlight the advantages of PCR-generated DNA templates as a rapid, efficient, and cost-effective alternative for mRNA synthesis, with broad applications in vaccine and therapeutic development.
体外转录（IVT）是一种广泛应用于基础研究和开发基于mRNA的疫苗和疗法的mRNA合成技术。IVT的效率主要取决于线性DNA模板的质量和完整性。传统的模板DNA制备方法包括在细菌中进行质粒增殖，然后进行酶线性化，这是一种劳动密集型和昂贵的方法。在这里，我们描述了一种无细胞、基于pcr的方法，用于生成高质量、高产率的线性DNA模板。我们在IVT效率、mRNA产量和mRNA- lnp（脂质纳米颗粒）疫苗的免疫原性方面广泛比较了基于pcr的方法与传统的基于质粒的方法。与质粒衍生的DNA相比，基于pcr的方法产生了更高数量的DNA模板和转录mRNA，同时保持了mRNA的质量和完整性。重要的是，两种方法生成的编码SARS-CoV-2刺突蛋白的mRNA-LNP疫苗在小鼠中引发了稳健且相似的免疫反应，两种模板方法之间没有观察到显著差异。我们的研究结果突出了pcr生成的DNA模板作为mRNA合成的一种快速、高效和经济的替代方法的优势，在疫苗和治疗开发中具有广泛的应用。
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引用次数: 0
Insights of Surface Enhancing Raman Spectroscopy for Biomedical Application 表面增强拉曼光谱在生物医学上的应用。
IF 4.3 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Methods
Pub Date : 2025-08-12 DOI: 10.1016/j.ymeth.2025.08.005
Neha Sharma , Anita Singh , Ratneshwar Kumar Ratnesh , Archana Adhana , Lalita Tyagi , Jay Singh
Raman spectroscopy is a powerful, non-invasive analytical technique that enables rapid identification of molecules based on their unique spectral fingerprints. Its sensitivity has been significantly enhanced through the use of metal nanoparticles in Surface-Enhanced Raman Spectroscopy (SERS), where molecules adsorbed on rough metallic surfaces or colloids produce Raman signals amplified by several orders of magnitude. This enhancement has opened new possibilities for molecular detection, particularly in surface chemistry and biomedical diagnostics.
In clinical applications, timely and accurate diagnosis is critical, yet conventional bioanalytical methods often require multiple biochemical tests, leading to delays that are especially problematic in emergency settings. SERS provides a promising alternative, offering high sensitivity, specificity, and rapid analysis with minimal sample preparation.
This review explores the integration of Raman spectroscopy—especially SERS—for both in vivo and ex vivo biomedical diagnostics. It covers sample preparation techniques, spectral data interpretation, and the correlation of Raman signals with disease-specific biomarkers. Special focus is given to the application of Raman-based methods in diagnosing brain disorders, various cancers, drug abuse, and COVID-19. Finally, the article discusses future prospects and challenges to guide the continued advancement of biomedical Raman technologies.
拉曼光谱是一种强大的非侵入性分析技术，可以根据分子独特的光谱指纹快速识别分子。通过在表面增强拉曼光谱（SERS）中使用金属纳米颗粒，其灵敏度得到了显着提高，其中分子吸附在粗糙的金属表面或胶体上产生拉曼信号，放大了几个数量级。这种增强为分子检测开辟了新的可能性，特别是在表面化学和生物医学诊断方面。在临床应用中，及时和准确的诊断至关重要，但传统的生物分析方法往往需要多次生化测试，导致延误，在紧急情况下尤其严重。SERS提供了一种很有前途的替代方案，提供高灵敏度，特异性和快速分析，只需最少的样品制备。本文综述了拉曼光谱-特别是sers -在体内和离体生物医学诊断中的应用。它涵盖了样品制备技术，光谱数据解释，以及拉曼信号与疾病特异性生物标志物的相关性。特别关注基于拉曼的方法在诊断脑部疾病、各种癌症、药物滥用和COVID-19中的应用。最后，文章讨论了未来的前景和挑战，以指导生物医学拉曼技术的持续发展。
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