Analytical biochemistry最新文献_第2页

An ATP detection system based on the enzyme reaction with biotin protein ligase 基于生物素蛋白连接酶反应的 ATP 检测系统。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-24 DOI: 10.1016/j.ab.2024.115698

Shinji Sueda, Satoshi Fujii

Adenosine triphosphate (ATP) is the energy currency of all living organisms and can be used as an indicator for cell proliferation and cytotoxicity. In the present work, we have developed a novel ATP detection system by combining the biotinylation reaction from archaeon Sulfolobus tokodaii with fluorescence resonance energy transfer (FRET). In biotinylation from S. tokodaii, an enzyme known as biotin protein ligase (BPL) forms a very stable complex with its product, biotinylated substrate protein (BCCP). Here, BPL and BCCP were fused to the fluorescent proteins Cerulean and Clover, respectively, and ATP detection was accomplished by monitoring the FRET signal between the two fluorescent proteins, since ATP is an essential component for biotinylation and the tight BPL-BCCP complex is formed only after biotinylation. Using this system, we have succeeded in detecting 5 nM of ATP by biotinylation reaction with 50 nM of each fusion protein. Our method has a characteristic that the signal does not decay for at least 2 h after the start of the reaction, unlike in the case of the luminescence-based assay with luciferase commonly used for the ATP detection. Thus, our system allows for ATP detection which is not significantly constrained by measurement timing.

三磷酸腺苷（ATP）是所有生物的能量货币，可用作细胞增殖和细胞毒性的指标。在本研究中，我们开发了一种新型的 ATP 检测系统，该系统将托克代磺酸古菌的生物素化反应与荧光共振能量转移（FRET）相结合。在托克达伊硫球菌的生物素化反应中，一种被称为生物素蛋白连接酶（BPL）的酶与其产物--生物素化底物蛋白（BCP）--形成非常稳定的复合物。在这里，BPL 和 BCCP 分别与荧光蛋白 Cerulean 和 Clover 融合，并通过监测两种荧光蛋白之间的 FRET 信号来实现 ATP 检测，因为 ATP 是生物素化的必要成分，而且只有在生物素化之后才会形成紧密的 BPL-BCCP 复合物。利用这一系统，我们成功地通过生物素化反应检测到了 5 nM 的 ATP 与 50 nM 的融合蛋白。我们的方法有一个特点，即信号在反应开始后至少 2 小时内不会衰减，这与常用于 ATP 检测的荧光素酶发光法不同。因此，我们的系统在进行 ATP 检测时不会受到测量时间的明显限制。

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引用次数: 0

Analytical and drug delivery strategies for short peptides: From manufacturing to market 短肽的分析和给药策略：从制造到上市。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-24 DOI: 10.1016/j.ab.2024.115699

Ashwini Chawathe , Vishal Ahire , Kshitiz Luthra , Bhumika Patil , Kalpna Garkhal , Nitish Sharma

In recent times, biopharmaceuticals have gained attention because of their tremendous potential to benefit millions of patients globally by treating widespread diseases such as cancer, diabetes and many rare diseases. Short peptides (SP), also termed as oligopeptides, are one such class of biopharmaceuticals, that are majorly involved in efficient functioning of biological systems. Peptide chains that are 2–20 amino acids long are considered as oligopeptides by researchers and are some of the functionally vital compounds with widespread applications including self-assembly material for drug delivery, targeting ligands for precise/specific targeting and other biological uses. Using functionalised biomacromolecules such as short chained peptides, helps in improving pharmacokinetic properties and biodistribution profile of the drug. Apart from this, functionalised SP are being employed as cell penetrating peptides and prodrug to specifically and selectively target tumor sites. In order to minimize any unwanted interaction and adverse effects, the stability and safety of SP should be ensured throughout its development from manufacturing to market. Formulation development and characterization strategies of these potential molecules are described in the following review along with various applications and details of marketed formulations.

近来，生物制药因其治疗癌症、糖尿病和许多罕见疾病等广泛疾病的巨大潜力而备受全球数百万患者的关注。短肽（SP），又称寡肽，就是这样一类生物制药，主要参与生物系统的有效运作。研究人员将长度为 2-20 个氨基酸的肽链视为寡肽，它们是一些具有重要功能的化合物，应用广泛，包括用于药物输送的自组装材料、用于精确/特异性靶向的靶向配体以及其他生物用途。使用短链肽等功能化生物大分子有助于改善药物的药代动力学特性和生物分布特征。除此之外，功能化 SP 还可用作细胞穿透肽和原药，以特异性和选择性地靶向肿瘤部位。为了最大限度地减少任何不必要的相互作用和不良反应，在从生产到上市的整个研发过程中，都应确保 SP 的稳定性和安全性。下文将介绍这些潜在分子的制剂开发和表征策略，以及各种应用和上市制剂的详细情况。

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引用次数: 0

Voltammetric-based immunosensing of Newcastle disease virus on polyethylene glycol-containing self-assembled monolayer modified gold electrode 含聚乙二醇的自组装单层修饰金电极上基于伏安法的新城疫病毒免疫传感。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-24 DOI: 10.1016/j.ab.2024.115700

Mohamad Farid Abd Muain , Amir Syahir Amir Hamzah , Suet Lin Chia , Khatijah Yusoff , Hong Ngee Lim , Ikeno Shinya , Asilah Ahmad Tajudin

A voltammetric immunosensor for the detection of Newcastle disease virus (NDV) has been developed by employing polyclonal antibody targeting NDV (anti-NDV) as a bioreceptor. Anti-NDV was immobilized on polyethylene glycol (PEG)-containing self-assembled monolayer (SAM) which was activated with N-(3-dimethylaminopropyl)-N′-ethylcarbodiimidehydrochloride (EDC) and N-hydroxy succinimide (NHS) coupling on screen-printed gold electrode (SPGE). The introduction of PEG-containing SAM on the SPGE allowed the bioreceptor to covalently bound to the electrode surface whilst still providing a hydrophilic layer on the electrode which is important to greatly reduce non-specific bindings. The bioreceptor functionalized electrode was then allowed to be incubated with NDV-spiked samples. The electrode surface modification with PEG-containing SAM, immobilization of anti-NDV as bioreceptor, up to the detection of NDV were characterized electrochemically through differential pulse voltammetry (DPV) analysis in [Fe(CN)₆]^3- as the redox probe. Decrement of anodic current peak (I_pa) of [Fe(CN)₆]^3- was seen as the concentration of NDV increased from 0.156 to 20 HA μL⁻¹ with the limit of detection (LoD) of 1.50 HA μL⁻¹ at 3σ m⁻¹. The detection of NDV in HA μL⁻¹ unit in this study would ease interlaboratory interpretation as it was the same unit used in hemagglutination (HA) assay of conventional NDV diagnosis. The specificity of anti-NDV used as bioreceptor towards NDV was confirmed through western blot analysis, whilst the selectivity of the bioreceptor-functionalized electrode has been tested with allantoic fluid as the negative control in which no apparent changes of anodic peak (I_pa) has been seen. This simple, fast, and less laborious electrochemical detection method could become an alternative to the conventional method for NDV detection.

利用针对新城疫病毒（NDV）的多克隆抗体（抗 NDV）作为生物受体，开发了一种用于检测新城疫病毒（NDV）的伏安免疫传感器。抗 NDV 被固定在含聚乙二醇（PEG）的自组装单层（SAM）上，SAM 经 N-(3-二甲基氨基丙基)-N'-乙基碳二亚胺盐酸盐（EDC）和 N-羟基琥珀酰亚胺（NHS）偶联后活化在丝网印刷金电极（SPGE）上。在 SPGE 上引入含 PEG 的 SAM 可使生物受体与电极表面共价结合，同时还能在电极上形成亲水层，这对大大减少非特异性结合非常重要。然后将生物受体功能化电极与添加了 NDV 的样品进行孵育。在[Fe(CN)6]3-作为氧化还原探针的条件下，通过差分脉冲伏安法（DPV）分析，对含有 PEG 的 SAM 的电极表面修饰、作为生物受体的抗 NDV 的固定以及 NDV 的检测进行了电化学表征。当 NDV 的浓度从 0.156 增至 20 HA μL-1 时，[Fe(CN)6]3- 的阳极电流峰值（Ipa）下降，3σ m-1 时的检测限（LoD）为 1.50 HA μL-1。本研究以 HA μL-1 为单位检测 NDV，这与传统 NDV 诊断的血凝（HA）检测所用的单位相同，因此便于实验室之间进行解释。抗 NDV 作为生物受体对 NDV 的特异性已通过 Western 印迹分析得到证实，而生物受体功能化电极的选择性已用尿囊液作为阴性对照进行了测试，结果显示阳极峰（Ipa）没有明显变化。这种简单、快速、省力的电化学检测方法可以替代传统的 NDV 检测方法。

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引用次数: 0

Research progress on Drug-Target Interactions in the last five years 过去五年药物与目标相互作用的研究进展。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-23 DOI: 10.1016/j.ab.2024.115691

Yun Zuo, Xubin Wu, Fei Ge, Hongjin Yan, Sirui Fei, Jingwen Liang, Zhaohong Deng

The identification of Drug-Target Interaction (DTI) is an important step in drug discovery and drug repositioning, and has high application value in multiple fields such as drug discovery, drug repositioning, and repurposing. However, the high cost of experimental validation limits its identification. In contrast, computation-based approaches are both economical and efficient. This review first synthesizes existing chemical genomic approaches, provides a comprehensive summary of prevalent databases for predicting DTIs, and categorizes the feature encodings from recent years. This is followed by an overview and brief description of the methods currently in use for predicting DTIs. The strengths and weaknesses of newly proposed prediction methods in the last five years (2020–2024), including those based on network representation learning and graph neural networks, are then discussed in detail, evaluating the performance of the different methods on a wide range of datasets. Finally, this review explores potential directions for future DTI research, emphasizing how to improve prediction accuracy and efficiency by combining big data and emerging computing technologies.

药物与靶点相互作用（DTI）的鉴定是药物发现和药物再定位的重要步骤，在药物发现、药物再定位和再利用等多个领域具有很高的应用价值。然而，高昂的实验验证成本限制了其鉴定。相比之下，基于计算的方法既经济又高效。本综述首先综述了现有的化学基因组学方法，全面总结了用于预测 DTI 的流行数据库，并对近年来的特征编码进行了分类。随后概述并简要介绍了目前使用的 DTI 预测方法。然后详细讨论了最近五年（2020-2024 年）新提出的预测方法（包括基于网络表示学习和图神经网络的预测方法）的优缺点，评估了不同方法在各种数据集上的性能。最后，本综述探讨了未来 DTI 研究的潜在方向，强调如何通过结合大数据和新兴计算技术来提高预测的准确性和效率。

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引用次数: 0

Detection of Salmonella by competitive ELISA of lipopolysaccharide secreted into the culture medium 通过对分泌到培养基中的脂多糖进行竞争性 ELISA 检测沙门氏菌。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-23 DOI: 10.1016/j.ab.2024.115695

Elena Kiseleva, Konstantin Mikhailopulo, Oleg Sviridov

Detection of Salmonella in food is topical due to known cases of salmonellosis epidemics. Immunochemical methods including ELISA are widely used for Salmonella detection. Traditionally, commercial ELISA kits are based on sandwich technique and detect lipopolysaccharide (LPS), which is considered to be the component of the outer membrane of Gram-negative bacteria. Our aim was elaboration of competitive ELISA test for Salmonella detection in food with improved parameters. It was shown that in the Salmonella culture after the standard sample preparation procedure LPS is present mainly outside cells as a component of outer membrane vesicles. Improved sample preparation procedure includes separation of bacteria from the medium and analysis of the medium, which increases analytical sensitivity. Immobilization of the bovine serum albumin (BSA)-LPS conjugate in microplate wells allows to obtain a more homogeneous coating than immobilization of LPS itself. Thus, we have developed test system for Salmonella detection in food by competitive ELISA of LPS secreted into the culture medium with the immobilized BSA-LPS conjugate and monoclonal antibodies (mAb) to LPS core in the liquid phase. New competitive ELISA test is high sensitive, give reproducible results, allows the detection of any Salmonella serotype and is important for the protection of human health.

由于已知的沙门氏菌病流行病例，检测食品中的沙门氏菌成为当务之急。包括 ELISA 在内的免疫化学方法被广泛用于检测沙门氏菌。传统上，商业 ELISA 试剂盒基于夹心技术，检测脂多糖（LPS），脂多糖被认为是革兰氏阴性细菌外膜的组成部分。我们的目标是研制一种竞争性 ELISA 检测试剂盒，用于检测食品中的沙门氏菌，并改进相关参数。结果表明，经过标准样品制备程序后，沙门氏菌培养物中的 LPS 主要作为外膜囊泡的成分存在于细胞外。改进后的样品制备程序包括从培养基中分离细菌和分析培养基，从而提高了分析灵敏度。在微孔板孔中固定牛血清白蛋白（BSA）-LPS 共轭物可获得比固定 LPS 本身更均匀的涂层。因此，我们开发了通过竞争性 ELISA 法检测食品中沙门氏菌的测试系统，用固定化的 BSA-LPS 结合物和液相中 LPS 核心的单克隆抗体（mAb）检测分泌到培养基中的 LPS。新的竞争性 ELISA 检测法灵敏度高，结果重复性好，可检测任何沙门氏菌血清型，对保护人类健康非常重要。

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引用次数: 0

Micro scale chromatography of human plasma proteins for nano LC-ESI-MS/MS 用于纳米 LC-ESI-MS/MS 的人体血浆蛋白质微尺度色谱法。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115694

Zhuo Zhen Chen, Jaimie Dufresne, Peter Bowden, Dominika Celej, Ming Miao, John G. Marshall

Organic precipitation of proteins with acetonitrile demonstrated complete protein recovery and improved chromatography of human plasma proteins. The separation of 25 μL of human plasma into 22 fractions on a QA SAX resin facilitated more effective protein discovery despite the limited sample size. Micro chromatography of plasma proteins over quaternary amine (QA) strong anion exchange (SAX) resins performed best, followed by diethylaminoethyl (DEAE), heparin (HEP), carboxymethyl cellulose (CMC), and propyl sulfate (PS) resins. Two independent statistical methods, Monte Carlo comparison with random MS/MS spectra and the rigorous X!TANDEM goodness of fit algorithm protein p-values corrected to false discovery rate q-values (q ≤ 0.01) agreed on at least 12,000 plasma proteins, each represented by at least three fully tryptic corrected peptide observations. There was qualitative agreement on 9393 protein/gene symbols between the linear quadrupole versus orbital ion trap but also quantitative agreement with a highly significant linear regression relationship between log observation frequency (F value 4,173, p-value 2.2e-16). The use of a QA resin showed nearly perfect replication of all the proteins that were also found using DEAE-, HEP-, CMC-, and PS-based chromatographic methods combined and together estimated the size of the size of the plasma proteome as ≥12,000 gene symbols.

用乙腈对蛋白质进行有机沉淀，可完全回收蛋白质并改进人血浆蛋白质的色谱分析。在 QA SAX 树脂上将 25 μL 人血浆分离成 22 个馏分，尽管样品量有限，但这有助于更有效地发现蛋白质。使用季胺（QA）强阴离子交换（SAX）树脂对血浆蛋白质进行微色谱分析的效果最好，其次是二乙胺基乙基（DEAE）、肝素（HEP）、羧甲基纤维素（CMC）和硫酸丙酯（PS）树脂。两种独立的统计方法：蒙特卡罗随机 MS/MS 图谱比较法和严格的 X!TANDEM 拟合度算法蛋白质 p 值校正为误差发现率 q 值（q≤ 0.01），对至少 12,000 种血浆蛋白质达成一致，每种蛋白质至少有三个完全胰蛋白酶校正的肽段观察结果。线性四极杆离子阱与轨道离子阱在 9393 个蛋白质/基因符号上存在定性一致，但也存在定量一致，观测频率对数之间存在高度显著的线性回归关系（F 值 4,173, p 值 2.2e-16）。使用 QA 树脂几乎完美地复制了使用 DEAE、HEP、CMC 和 PS 色谱方法发现的所有蛋白质，并估算出血浆蛋白质组的大小≥12,000 个基因符号。

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引用次数: 0

Uracil base PCR implemented for reliable DNA walking 实施尿嘧啶碱基 PCR，实现可靠的 DNA 走查。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115697

Rongrong Wang , Yinwei Gu , Hong Chen , Bingkun Tian , Haixing Li

PCR-based DNA walking is of efficacy for capturing unknown flanking genomic sequences. Here, an uracil base PCR (UB-PCR) with satisfying specificity has been devised for DNA walking. Primary UB-PCR replaces thymine base with uracil base, resulting in a primary PCR product composed of U-DNAs. A single-primer (primary nested sequence-specific primer) single-cycle amplification, using the four normal bases (adenine, thymine, cytosine, and guanine) as substrate, is then performed on the primary PCR product. Clearly, only those U-DNAs, ended by the primary nested sequence-specific primer at least at one side, will produce the corresponding normal single strands. Next, the single-cycle product undergoes uracil-DNA glycosylase treatment to destroy the U-DNAs, while the normal single strands are unaffected. Afterward, secondary even tertiary PCR is performed to exclusively enrich the target product. The feasibility of UB-PCR has been checked by obtaining unknown sequences bordering the three selected genetic sites.

基于 PCR 的 DNA 走查对于捕获未知的侧翼基因组序列非常有效。在此，我们设计了一种具有良好特异性的尿嘧啶碱基 PCR（UB-PCR）来进行 DNA 扩增。初级 UB-PCR 将胸腺嘧啶碱基替换为尿嘧啶碱基，从而产生由 U-DNA 组成的初级 PCR 产物。然后，以四个正常碱基（腺嘌呤、胸腺嘧啶、胞嘧啶和鸟嘌呤）为底物，对主 PCR 产物进行单引物（主嵌套序列特异性引物）单循环扩增。显然，只有那些至少一边以主嵌套序列特异性引物为末端的 U-DNA 才能产生相应的正常单链。接下来，单循环产物经过尿嘧啶-DNA 糖基化酶处理，以破坏 U-DNA，而正常的单链则不受影响。之后，进行二次甚至三次 PCR，以专门富集目标产物。通过获得与三个选定基因位点相邻的未知序列，检验了 UB-PCR 的可行性。

引用次数: 0

Electrochemical biosensor based on copper sulfide/reduced graphene oxide/glucose oxidase construct for glucose detection 基于硫化铜/还原氧化石墨烯/葡萄糖氧化酶构建的电化学生物传感器用于葡萄糖检测。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-22 DOI: 10.1016/j.ab.2024.115696

Yuchen Zhang , Jiangnan Chen , Huifang Wang , Xianghua Gao , Baolong Niu , Wenfeng Li , Hong Wang

Due to the current increase in the number of people suffering from diabetes worldwide, how to monitor the blood glucose level in the human body has become an urgent problem to be solved nowadays. The electrochemical sensor method can be used for real-time glucose monitoring due to its advantages of real-time monitoring capability and high sensitivity. Reduced graphene oxide (rGO) has great potential for application in the field of sensors due to its advantages of large specific surface area, high stability, and good electrical and thermal conductivity. Meanwhile, the synergistic effect between two-dimensional transition metal sulfides and graphene can improve the electrochemical performance of materials due to their similar mechanical flexibility and strength. This article uses flake graphite, copper sulfate, and glucose oxidase (GOx) as raw materials to prepare CuS/rGO/GOx/GCE electrodes, and explores the performance of electrode electrocatalysis for glucose. The results showed that the prepared sensor was characterized by a low detection limit (1.75 nM) and a wide linear range (0.1–100 mM) for glucose detection, displaying a good overall detection performance, and its sensing mechanism and dynamic process were also investigated. In addition, the sensor has outstanding selectivity, anti-interference, repeatability, reproducibility and practicality.

由于目前全球糖尿病患者人数的增加，如何监测人体血糖水平已成为当前亟待解决的问题。电化学传感器方法具有实时监测能力强、灵敏度高等优点，可用于葡萄糖的实时监测。还原氧化石墨烯（rGO）具有比表面积大、稳定性高、导电导热性好等优点，在传感器领域具有很大的应用潜力。同时，由于二维过渡金属硫化物与石墨烯具有相似的机械柔韧性和强度，二者之间的协同效应可以提高材料的电化学性能。本文以鳞片石墨、硫酸铜和葡萄糖氧化酶（GOx）为原料制备了 CuS/rGO/GOx/GCE 电极，并探讨了电极对葡萄糖的电催化性能。结果表明，所制备的传感器对葡萄糖的检测具有检出限低（1.75 nM）、线性范围宽（0.1-100 mM）的特点，显示出良好的整体检测性能，并对其传感机理和动态过程进行了研究。此外，该传感器还具有突出的选择性、抗干扰性、重复性、再现性和实用性。

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引用次数: 0

Impact of Lectin biotinylation for surface plasmon resonance and enzyme-linked Lectin assays for protein glycosylation 表面等离子共振和酶联凝集素检测蛋白质糖基化的凝集素生物素化的影响。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-19 DOI: 10.1016/j.ab.2024.115693

Benjamin Serafin , Amine Kamen , Gregory De Crescenzo , Olivier Henry

Lectins are widely employed for the assessment of protein glycosylation as their carbohydrate binding specificities have been well characterized. In glycosylation assays, lectins are often conjugated with biotin tags, which interact with streptavidin to functionalize biosensing surfaces or recruit signal generating molecules, depending on the assay configuration. We here demonstrate that a high degree of biotin conjugation can limit total capture to streptavidin functionalized SPR surfaces due to multipoint binding, and can additionally bias the reported kinetic evaluations when measuring the interaction between lectins and glycoproteins by SPR. For microplate assays using different configurations, high biotinylation ratios can effectively amplify the signal obtained when using Streptavidin conjugates for detection, in some cases significantly lowering the limit of detection. The cumulative results express the importance of customizing the ligand biotinylation ratios for different assay configurations, as commercially obtained pre-biotinylated lectins are not necessarily optimized for different assay configurations.

凝集素被广泛用于评估蛋白质糖基化，因为它们与碳水化合物结合的特异性已经得到了很好的表征。在糖基化检测中，凝集素通常与生物素标签连接，生物素标签与链霉亲和素相互作用，使生物传感表面功能化，或根据检测配置吸附信号生成分子。我们在此证明，由于多点结合，高度的生物素共轭会限制链霉亲和素功能化 SPR 表面的总捕获量，而且在通过 SPR 测量凝集素和糖蛋白之间的相互作用时，还会使报告的动力学评估出现偏差。对于使用不同配置的微孔板检测，当使用链霉亲和素共轭物进行检测时，高生物素化比率可有效放大获得的信号，在某些情况下可显著降低检测限。累积的结果表明了针对不同的检测配置定制配体生物素化比率的重要性，因为市售的预生物素化凝集素并不一定针对不同的检测配置进行了优化。

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引用次数: 0

The downstream purification of bispecific antibodies 双特异性抗体的下游纯化。

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry

Pub Date : 2024-10-18 DOI: 10.1016/j.ab.2024.115692

Qian Li , Hongyang Zhao , Xiaoying Liang, Qingquan He, Zicheng Wang, Guohong Qin, GuoZhu Li, Dan Xu

Bispecific antibodies, a class of therapeutic antibodies, can simultaneously bind to two distinct targets. Compared with monospecific antibodies, bispecific antibodies offer advantages, including superior efficacy and reduced side effects. However, because of their structural complexity, the purification of bispecific antibodies is highly challenging. The purification process must strike a delicate balance between purity and productivity, eliminating a broad spectrum of contaminants, including product-related and process-related impurities, while also maximizing the yield wherever feasible. This review systematically describes the strategies for bispecific antibody capture, the elimination of product-related impurities, and the mitigation of the formation of process-related impurities, thereby, providing guidance for the development of downstream purification processes for bispecific antibodies.

双特异性抗体是一类治疗性抗体，可同时与两个不同的靶点结合。与单特异性抗体相比，双特异性抗体具有疗效更佳、副作用更小等优点。然而，由于其结构复杂，双特异性抗体的纯化极具挑战性。纯化过程必须在纯度和产量之间取得微妙的平衡，既要剔除各种杂质，包括与产品和工艺相关的杂质，又要尽可能提高产量。本综述系统地阐述了双特异性抗体捕获、消除产品相关杂质和减少工艺相关杂质形成的策略，从而为双特异性抗体下游纯化工艺的开发提供指导。

引用次数: 0