评价自身抗体联合检测在诊断自身免疫性疾病中的应用

目的探讨检测抗核抗体（ANA）核型与抗可提取性核抗原（ENA）抗体及抗双链DNA（ds,DNA）抗体之间的相关性,提高对自身免疫性疾病（AID）的诊断及鉴别诊断参考价值。方法资料来源门诊及住院的AID病例血清标本进行检测,ANA检测采用问接免疫荧光法（IIF）,抗ENA抗体检测采用欧蒙斑点法（抗nRNP／SIn、Sm、SSA／Ro、SSB／Lo、Scl-70和Jo—1）六项抗体及抗（ds—DNA）抗体采用ELISA法检测。结果在98例ANA阳性病例血清标本中核型分布为：细胞核均质型（36．74％）,细胞核颗粒型（41．84％）,细胞核仁型（11．22％）,细胞核膜型（1．02％）,细胞核着丝点型（1．02％）;重叠核型中细胞核均质型／细胞核颗粒型（2．04％）,细胞核均质型／细胞浆颗粒型（6．12％）等。98例ANA与ENA同时阳性血清标本中42例抗双链DNA抗体阳性（42．86％）。36例均质型阳性样本中抗SSA／Ro抗体（24．48％）,抗SSB／Lo抗体（6．12％）,抗nRNP／Sm抗体（6．12％）。41例细胞核颗粒型阳性样本抗nRNP／Sm抗体（19．39％）,SSAfRo抗体（15．31％）,抗SSB／Lo抗体（6．12％）。ANA阳性可出现一种或多种自身抗体。结论ANA常见核型有：细胞核颗粒型,细胞核均质型及细胞核仁型。ANA均质型核型检出的抗ds-DNA抗体为主,且抗ds—DNA抗体含量与荧光强度成正比。ANA核型与抗ENA抗体二者之间有一定的相对应性。ANA及核型、抗ENA抗体和抗ds—DNA抗体的联合检测可明确诊断AID,而且利于判断患者血清中各种自身抗体与临床表现的关系。     

基于UPLC-Q-TOF-MS的双特异性抗体关键表征方法研究

目的 建立Y-Body^®型双特异性抗体一级结构的超高效液相色谱-三重四级杆/飞行时间质谱联用技术(UPLC-Q-TOF-MS)研究方法。方法 以Y-Body^®型双特异性抗体M808为研究对象,基于UPLC-Q-TOF-MS检测完整抗体及各亚基去糖基化处理前后的相对分子质量、肽图(氨基酸序列),并通过Unify软件进行解析。结果 完整抗体去糖基化处理前后的相对分子质量分别为128417.6605,125040.7210;轻链去糖基化处理前后的相对分子质量分别为23447.2322,23445.5608;重链去糖基化处理前后的相对分子质量分别为50804.7732,49201.6051;单链去糖基化处理前后的相对分子质量分别为54189.9224,52415.7644;其氨基酸序列与理论序列基本一致,肽图的互补决定区覆盖率为100%;推测N-糖基化位点位于重链及单链Fc端,糖型以G1F和G2F为主。结论 通过UPLC-Q-TOF-MS及Unify软件可有效表征双特异性抗体的一级结构,为其质量标准的制定提供依据。     

四氢异喹啉类生物碱的生物活性多样性及其作用机制

四氢异喹啉类生物碱在自然界中分布广泛, 有些药物在临床上已广泛应用。随着对四氢异喹啉类生物碱研究的深入, 越来越多新的生物活性、新的作用机制和靶点被发现和揭示, 此类生物碱生物活性的多样性受到人们广泛关注。本文综述了近五年来四氢异喹啉类化合物在抗肿瘤、抗菌、抗病毒、抗炎、抗凝血、支气管扩张及中枢神经系统作用等方面的活性及机制研究的新进展, 旨在为四氢异喹啉类生物碱的活性研究提供思路和启示, 为寻找先导化合物、合理设计药物分子提供依据。     

Recent advances in the generation of chemical diversity libraries

In recent years, screening in combination with a diverse compound collection has become a powerful method for discovering leads for the ever-increasing number of new biologically active peptides, proteins, receptors, and enzymes discovered continually. As a result, the rapid generation and screening of compound libraries (collections) have recently become important major tools in the search for novel lead structures. Diverse collections of compounds have been acquired by many strategies; these include (1) natural products from plants, fermentation, marine organisms, insect toxins, and ethnic pharmacotherapies; (2) recombinant randomized peptide libraries (often referred to as biological diversity); (3) multiple peptide synthesis; and (4) non-peptidic synthetic libraries. The present review provides an overview of the recent advances in the field of peptide and non-peptidic synthetic libraries. The progress made thus far is broadly divided into two categories: (1) Amide based libraries. These libraries share the concepts of the peptide library strategies; much of the referenced work thus refers to peptides, reflecting the bias of the literature to date. (2) Non-amide based libraries. This promising technology combines solid phase synthesis with classical organic synthesis to provide large numbers of compounds with desirable bioavailability and pharmacokinetics for screening. The basic premise behind the second approach is that the high affinity ligands, when identified, will be much more likely to become useful therapeutic agents than the compounds discovered from amide based libraries. Synthesizing small heterocyclic ring systems that use ligands of diverse biological activity via combinatorial strategies is a fast developing branch of medicinal chemistry. We are at an early state in the development of combinatorial chemistry. However, this dramatic convergence of technologies represents a fundamental advance in medicinal chemistry and promises to play a major role in future drug discovery efforts. © 1994 Wiley-Less, Inc.     

Neutralizing Monoclonal Antibody,mAb 10D8, Is an Effective Detoxicant against Abrin-a Both In Vitro and In Vivo

Abrin is a types II ribosome-inactivating protein (RIP) isolated from Abrus precatorious seeds, which comprises a catalytically active A chain and a lectin-like B chain linked by a disulfide bond. Four isotoxins of abrin have been reported with similar amino-acid composition but different cytotoxicity, of which abrin-a is the most potent toxin. High lethality and easy availability make abrin a potential bioterrorism agent. However, there are no antidotes available for managing abrin poisoning, and treatment is only symptomatic. Currently, neutralizing antibodies remain the most effective therapy against biotoxin poisoning. In this study, we prepared, identified, and acquired a high-affinity neutralizing monoclonal antibody (mAb) 10D8 with a potent pre- and post-exposure protective effect against cytotoxicity and animal toxicity induced by abrin-a or abrin crude extract. The mAb 10D8 could rescue the mouse injected intraperitoneally with a 25 × LD₅₀ dose of abrin-a from lethality and prevent tissue damages. Results indicated that 10D8 does not prevent the binding and internalization of abrin-a to cells but inhibits the enzymatic activity of abrin-a and reduces protein synthesis inhibition of cells. The high affinity, good specificity, and potent antitoxic efficiency of 10D8 make it a promising candidate for therapeutic antibodies against abrin.     

Genetic diversity: applications of evolutionary algorithms to combinatorial library design

Combinatorial chemistry and other high-throughput techniques continue to affect the drug discovery process in a profound manner. This review examines the impact of combinatorial chemistry upon the discipline of computer-aided molecular design and describes the methods that are being developed to enable researchers to quantify molecular diversity and to design efficient combinatorial libraries. In particular, the application of optimisation algorithms based on evolutionary principles to these problems is examined and illustrated with examples from the literature.     

Salt-dependent conformational diversity of alternating poly (Glu-Leu)

Alternating poly(Glu-Leu) was synthesized by the condensation of the corresponding dipeptide p-nitrophenyl ester at high concentration. It exhibits a random coil structure in pure water at neutral pH. Addition of monovalent cations, such as NH⁺₄ to a final 0.1 M solution, induces a transition to a water soluble β-structure. The salt effect is quite selective since no transition was observed with Li⁺, Na⁺ or Cs⁺ ions. Addition of 0.5 equiv. of calcium, cobalt or manganese chlorides per glutamyl residue induces similar coil to β-sheet transitions. No polymer precipitation was observed at these very low salt concentrations. Addition of 0.5 equiv. of Cu²⁺ or 0.15 equiv. of Fe³⁺ induces a coil to α-helix transition. Molecular modeling has been used to understand tentatively the main factors controlling the different conformations observed with the various metal ions. © Munksgaard 1997.     

单克隆抗体药物进展

单克隆抗体药物特异性高、结构与性质均一稳定,其制备技术日益完善,临床应用越来越广泛,已有18种产品上市并用于人类疾病的诊断和治疗,100多种单克隆抗体药物进入临床研究阶段;单克隆抗体药物研究在生物技术药物中占有突出位置。文章综述了其国内外研究开发情况。     

高通量DNA测序技术在抗体新药研发中的应用

自2005年首次报道了一种基于微乳液PCR技术的高通量DNA测序技术 (high-throughput DNA sequencing technology) 以来, 高通量DNA测序平台已经发展为基因组和各种基因文库序列检测的强大工具。大容量的抗体基因库是目前获得抗体新药的基础, 高通量DNA测序技术为从海量的抗体基因库中快速发现功能抗体分子提供了可能。本文就近几年高通量DNA测序技术在抗体基因库的多样性分析, 抗体CDR3区的高通量测序、频率分析、功能基因发现及各种展示技术与高通量DNA测序技术的对接应用等方面进行了综述, 以期为抗体新药的研发提供一条新的技术路线。     

抗体药物在肿瘤治疗中的应用与进展

单克隆抗体药物以其特异性高、亲和力强、血清半衰期长等优点在肿瘤治疗领域得到广泛应用。基于母体单克隆抗体而设计的双特异性抗体、抗体片段、抗体药物偶联物等将抗体类药物的开发推至新的高潮。通过对抗体药物在肿瘤治疗领域的应用和最新进展、抗体药物的结构和作用机制进行介绍,并对肿瘤治疗抗体药物上市与临床开发现状进行总结,以期为抗体药物在肿瘤治疗领域的未来发展提供参考。     

双特异性抗体的药动学研究进展

双特异性抗体（BsAb）是指能同时结合相同或不同分子上的2个不同抗原表位的基因工程抗体。目前处于临床试验阶段的BsAb多达百余种,其适应证也涵盖肿瘤、自身免疫系统疾病等多个领域,有着广阔的应用前景。生物技术的高速发展提供了多种构建BsAb的模式,而不同构建模式的BsAb在体内的吸收、分布、代谢、外排的动力学行为和特征不同。深入研究BsAb药动学行为及其影响因素,对BsAb的合理设计有重要指导意义。通过从BsAb的构建模式出发,同时详细比较不同构建模式的BsAb在体内的药动学表现的差异,以期为BsAb药物设计和临床试验进一步提供参考。     

Increased diversity of libraries from libraries: chemoinformatic analysis of bis-diazacyclic libraries

Combinatorial libraries continue to play a key role in drug discovery. To increase structural diversity, several experimental methods have been developed. However, limited efforts have been performed so far to quantify the diversity of the broadly used diversity-oriented synthetic libraries. Herein, we report a comprehensive characterization of 15 bis-diazacyclic combinatorial libraries obtained through libraries from libraries, which is a diversity-oriented synthetic approach. Using MACCS keys, radial and different pharmacophoric fingerprints as well as six molecular properties, it was demonstrated the increased structural and property diversity of the libraries from libraries over the individual libraries. Comparison of the libraries to existing drugs, NCI diversity, and the Molecular Libraries Small Molecule Repository revealed the structural uniqueness of the combinatorial libraries (mean similarity <0.5 for any fingerprint representation). In particular, bis-cyclic thiourea libraries were the most structurally dissimilar to drugs retaining drug-like character in property space. This study represents the first comprehensive quantification of the diversity of libraries from libraries providing a solid quantitative approach to compare and contrast the diversity of diversity-oriented synthetic libraries with existing drugs or any other compound collection.     

ACA、AsAb与不孕及反复自然流产的关系

目的 探讨抗心磷脂抗体(ACA)、抗精子抗体(AsAb)与不孕、反复自然流产的关系.方法 采用酶联免疫吸附法(ELISA)对124例不孕患者(不孕组)、182例反复自然流产(流产组)及68例无流产史且已分娩的健康妇女(对照组)血清ACA及AsAb进行测定分析并比较.结果 不孕组ACA阳性率为45.16%,AsAb阳性率...     

单克隆抗体纯化研究进展

综述了单克隆抗体的纯化技术,讨论了各单元操作的关键问题和解决方法,并重点介绍了模拟移动床色谱、双水相萃取、膜色谱等操作简单、能连续生产、低成本的纯化技术。     

哺乳动物细胞表面展示人源HIV特异性全长抗体库的构建

目的构建人源的HIV特异性全长抗体基因库并获得可稳定展示全长抗体的哺乳动物细胞库。方法提取HIV患者外周血淋巴细胞的总RNA,采用RT-PCR的方法扩增抗体重链可变区(VH)基因,插入哺乳动物细胞表达载体pDGB4,转化感受态大肠杆菌TOPO-10,构建抗体基因库,随后将抗体库的质粒DNA稳定转染FCHO细胞,获得可稳定展示全长抗体的细胞库,用流式细胞仪分析全长抗体在FCHO细胞表面的表达。结果本研究以少量外周血淋巴细胞RNA为来源,构建了库容量为7.77×104的全长抗体基因库;将基因库的质粒DNA稳定转染FCHO细胞后,获得了可稳定展示全长抗体的细胞库;经流式细胞仪分析,有67%的细胞表面可表达可被检测到的全长抗体。结论本研究构建了人源的HIV特异性全长抗体基因库并获得可稳定展示全长抗体的哺乳动物细胞库,为治疗HIV感染的特异性抗体的筛选打下了基础。     

胰岛素致外源性胰岛素抗体综合征患者的临床特点

目的 探讨胰岛素导致外源性胰岛素抗体综合征(EIAS)患者的临床特点及随访24周临床转归情况.方法 纳入2015年1月-2018年6月诊断的使用胰岛素治疗后出现EIAS的患者16例.经调整药物治疗后于第12周、24周随访,观察空腹血糖(FBG)、糖化血红蛋白(HbA1c)、空腹胰岛素(FINS)、餐后2h胰岛素(2hI...     

The problems of memory in aging: Molecular aspects

Mandel, P. :. The problems of memory in aging: Molecular aspects. Drug Dev. Res. 2: 431–432, 1982. It is generally accepted that molecular mechanisms are involved in induction of memory engrams. Thus one may expect that aging, which alters brain metabolism, will also alter memory. The free-radical theory and the cross-linking theory of aging seem to be the most tempting for speculating about memory alterations. The loss of neurons and of their circuits, which probably plays a fundamental role in memory alternation, may result from free-radical effects that alter DNA, induce an increase of repair phenomena, and thus favor mistakes in the repair of DNA.     

静注人免疫球蛋白对新生儿乙肝核心抗体的影响

目的:探讨新生儿使用静注人免疫球蛋白(IVIG)后乙肝核心抗体(HBcAb)的变化。方法:采用回顾性研究方法,收集我院2019-2020年使用IVIG的住院新生儿病历,分析使用IVIG前后患儿HBcAb的变化。结果:共收集43例患儿,其中5例使用IVIG前后HBcAb均呈阳性,10例使用IVIG前后HBcAb均呈阴性,18例使用IVIG后HBcAb由阴性转为阳性,10例使用IVIG后因未进行乙肝5项复测未知HBcAb的变化。结论:输注IVIG会引起新生儿HBcAb水平升高,提示对输注IVIG的患儿进行抗体被动转移的排除具有重要意义。     

Antibody therapy of lymphoma

The availability of rituximab and the possible imminent availability of two new radiolabelled monoclonal anti-CD20 antibodies (Yttrium-90 (⁹⁰Y)-ibritumomab and Iodine-131(¹³¹I)-tositumomab) have captured much attention in the treatment of lymphoma. The chimeric monoclonal anti-CD20 antibody, rituximab has truly heralded a new era for the treatment of lymphoma and human malignancies. The full potential of antibody-based therapy to improve the outcome in patients with B-cell non-Hodgkin’s lymphoma has yet to be defined, but recent data suggests that the combination of chemotherapy plus rituximab may significantly improve outcome for patients with aggressive lymphoma over chemotherapy alone. Highly promising data are also emerging for the use of rituximab in combination with chemotherapy in other types of lymphoma. New advances in antibody therapy, driven by new technologies and defining novel antigen targets, offer the promise of more effective tumour specific therapies. Combinations of antibodies, either conjugated with radioisotopes or unlabelled, used with chemotherapy are likely to provide definitive advances in the treatment of lymphoma in the immediate future.