Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp简

A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2–4F, 7–3G, and 11–11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC50) and percentage of cross-reactivity, respectively. MAb 2–4F exhibited the highest sensitivity, with an IC₅₀ of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2–4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively.     

单克隆抗体研究进展——不同表达系统介绍与比较

单克隆抗体的高度特异性使其在包括细胞生物学、基础医学、临床诊断等领域得到广泛应用,而各种表达系统的开发进一步促进了单克隆抗体及其衍生物的发展.对目前用于生产单克隆抗体的主要表达系统进行综述,同时比较各系统的特点和优缺点.     

单克隆抗体在肿瘤治疗中的应用

随着一些单克隆抗体获准被应用于临床,单抗在肿瘤治疗中的运用得到了快速发展。其中代表性的有抗血管内皮生长因子的单抗bevacizumab(Avastin),和抗表皮生长因子的cetuximab(Erbitux)。结合常规的化疗,bevacizumab能够显著延长结肠癌、乳腺癌及肺癌患者的生命。Cetuximab无论与化疗联合使用或单独使用对于对化疗有耐药性的结肠癌直肠癌均有显著的临床意义。     

抗体药物的研究进展

随着人们对抗体的结构与功能的了解不断深入,抗体不仅在抵抗外来生物物质方面有着重要作用,而且利用它本身的特点来防治疾病更是人们的研究热点。特别是单克隆抗体技术的问世,使得研究和生产单抗药物成为现实。本文就单抗药物的历史、现状、存在的问题及未来展望做一简要综述。     

Nicotinamide <Emphasis Type="Italic">N</Emphasis>-methyltransferase protein expression in renal cell cancer

Objective: To understand the function of nicotinamide N-methyltransferase (NNMT) protein as tumor biomarker in renal carcinoma. Methods: Recombinant NNMT protein was used to prepare monoclonal antibodies by hybridoma technique. The diagnostic and prognostic function of NNMT protein in renal carcinoma was evaluated by analyzing 74 renal cancer tissues through immunohistochemical staining for NNMT by using the prepared antibodies. Results: Two hybridomas named 2F8 and 1E7 stably secreting the monoclonal antib...     

体积排阻色谱法分离单克隆抗体的条件优化

探讨了流动相条件及流速对体积排阻色谱分离抗EGF单克隆抗体的影响.结果表明:选择100mmol/L磷酸钠-150mmol/L氯化钠(pH=7.0)为流动相,设定流速为0.6mL/min时,抗EGF单克隆抗体在体积排阻色谱柱上的分离能获得最佳柱效.     

口臭菌分离及卵黄抗体制备综合性实验

为了激发学生对实验的兴趣,培养学生的综合能力,设计了一个综合性实验。该实验首先分离和培养了口臭菌,并对其进行形态观察,然后制备成疫苗,通过免疫、分离提取和鉴定,获得了卵黄抗体IgY。该实验涉及多个学科的多种类型实验,以期提高学生学习的时效性及有利于对学生综合能力的培养。     

Anti-CD69 monoclonal antibody treatment inhibits airway inflammation in a mouse model of asthma

Objective: Airway inflammation and airway hyper-responsiveness (AHR) are principle pathological manifestations of asthma. Cluster of differentiation 69 (CD69) is a well-known co-stimulatory factor associated with the activation, proliferation as well as apoptosis of immune cells. This study aims to examine the effect of anti-CD69 monoclonal antibody (mAb) on the pathophysiology of a mouse model of asthma. Methods: A murine model of ovalbumin (OVA)-induced allergic airway inflammation was used in this study. Briefly, mice were injected with 20 μg chicken OVA intraperitoneally on Days 0 and 14, followed by aerosol provocation with 1% (0.01 g/ml) OVA on Days 24, 25, and 26. Anti-CD69 mAb or isotype IgG was injected intraperitoneally after OVA challenge; dexamethasone (DXM) was administrated either before or after OVA challenge. AHR, mucus production, and eosinophil infiltration in the peribronchial area were examined. The levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-5 (IL-5) in bronchoalveolar lavage fluid (BALF) were also assayed as indices of airway inflammation on Day 28 following OVA injection. Results: Pretreatment with DXM together with anti-CD69 mAb treatment after OVA provocation completely inhibited AHR, eosinophil infiltration and mucus overproduction, and significantly reduced BALF IL-5. However, treatment with DXM alone after OVA challenge only partially inhibited AHR, eosinophil infiltration and mucus overproduction, and did not diminish BALF IL-5. Treatment with either DXM or anti-CD69 mAb did not alter the concentration of BALF GM-CSF. Conclusions: Anti-CD69 mAb treatment inhibits established airway inflammation as effectively as DXM pretreatment. This study provides a potential alternative therapeutic opportunity for the clinical management of asthma and its exacerbation.     

一种快速检测水稻条纹病毒的胶体金免疫层析试纸条的研制（英文）

目的:制备一种检测水稻植物和传毒介体灰飞虱中水稻条纹病毒(RSV)的胶体金免疫层析试纸条,为水稻条纹叶枯病的田间调查和检测以及预测预警提供快速、实用的检测试剂。创新点:利用制备的RSV单克隆抗体,首次制备了能在5～10 min内快速、准确、灵敏、特异地检测水稻植物和灰飞虱体内RSV的胶体金免疫层析试纸条。方法:以差速离心方法提纯的RSV病毒粒子作为免疫原免疫BALB/c小鼠,通过杂交瘤技术获得了高度特异和灵敏的RSV单克隆抗体。采用柠檬酸钠还原氯金酸的方法制备胶体金并标记一个RSV单克隆抗体,另一个RSV单克隆抗体和羊抗鼠抗体分别包被到硝酸纤维素膜的检测线和质控线,将吸水滤纸制成的样品垫、RSV免疫胶体金垫、结合有RSV单抗和羊抗鼠抗体的硝酸纤维素膜和吸水纸依次粘贴到聚氯乙烯(PVC)胶板上研制检测水稻植物和传毒介体灰飞虱中RSV的胶体金免疫层析试纸条。对田间样品进行RSV检测,分析试纸条检测RSV的有效性。结论:利用杂交瘤技术获得了2株RSV单克隆抗体(16E6和11C1),胶体金标记16E6单克隆抗体包被在聚酯膜制成的结合垫上,以11C1单克隆抗体和羊抗鼠抗体分别包被到硝酸纤维素膜的检测线和质控线,制成能在5～10 min内快速、特异、灵敏、准确地检测田间水稻及单头灰飞虱样品中RSV的胶体金免疫层析试纸条。试纸条检测感染RSV的水稻植株和灰飞虱呈阳性反应,而检测健康水稻和感染其它5种常见水稻病毒的植株及其传毒介体呈阴性反应,且试纸条检测RSV感染水稻植物组织的灵敏度达到1:20 480倍稀释(w/v, g/mL),检测携带RSV单头灰飞虱的灵敏度达到1:2560倍稀释(单头灰飞虱/μL)。田间样品检测结果发现,试纸条检测结果与反转录聚合酶链反应(RT-PCR)的检测结果一致,表明制备的试纸条可有效地用于田间RSV的检测,从而为水稻条纹叶枯病的诊断、监测预警、抗病育种、流行病学研究及科学防治提供技术和物质支撑。     

嵌合抗原受体T细胞在治疗难治/复发多发性骨髓瘤中的新进展（英文）

多发性骨髓瘤被认为是一种无法治愈的血液系统恶性疾病,其特征为恶性浆细胞的克隆性增殖。尽管在过去的几十年中,自体干细胞移植(ASCT)的应用及新型药物(蛋白酶体抑制剂和免疫调节药)的问世,将患者的中位生存时间由原来的4年提高到了8年,但复发与难治仍然是多发性骨髓瘤疾病进程中难以逾越的鸿沟。为了获得长期持续的缓解,免疫治疗开始在多发性骨髓瘤中崭露头角,其中嵌合抗原受体(CAR)T细胞治疗就是最有潜力的一颗新星。通过在基因层面改造患者自己的T细胞,使T细胞表达一种特定的受体(人造的融合蛋白),该受体可以识别并结合肿瘤相关抗原,并活化T细胞启动后续的杀伤过程。Tisagenlecleucel和Axicabtagene是两个针对CD19抗原的CAR T产品,用于治疗B细胞来源的急性淋巴细胞白血病(B-ALL)和弥漫大B细胞淋巴瘤(DLBCL),并于2017年被美国食品药品监督管理局(FDA)批准。这两个产品的发展极大推动了B细胞来源的恶性血液系统疾病的治疗,并刷新了对于传统治疗的认知。基于之前CAR T治疗的成功经验,寻找如CD19一样的特定靶点能为CAR T治疗多发性骨髓瘤打下基础。本综述介绍了数个在骨髓瘤细胞上的肿瘤靶抗原,如B细胞成熟抗原(BCMA)和CD38。这些针对抗原的CAR T治疗有些还在实验室阶段,而有些已经进入了3期的临床试验,很有可能成为下一个被批准的CAR T产品。另外,本综述也介绍了在CAR T治疗中出现的毒副反应以及相应的管理和处理方法。     

Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine

A protein conjugate of streptomycin (streptomycin-bovine serum albumin (BSA) conjugate) was prepared and used as immunogen to produce monoclonal antibodies (MAb). One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentration (IC50) value of 4.65 ng/ml and the IC20 value of 0.21 ng/ml in phosphate buffered saline (PBS). At optimum conditions, an indirect competitive enzyme-linked immunosorbent assay (ELISA) and a co...     

Preparation of high-affinity rabbit monoclonal antibodies for ciprofloxacin and development of an indirect competitive ELISA for residues in milk

A convenient competitive enzyme-linked immunosorbent assay (ELISA) for ciprofloxacin (CPFX) was developed by using rabbit monoclonal antibodies (RabMAbs) against a hapten-protein conjugate of CPFX-bovine serum albumin (BSA). The indirect competitive ELISA of CPFX had a concentration at 50% inhibition (IC₅₀) of 1.47 ng/ml and a limit of detection (LOD) of 0.095 ng/ml. The mAb exhibited some cross-reactivity, however, not so high with enrofloxacin (28.8%), ofloxacin (13.1%), norfloxacin (11.0%), fleroxacin (22.6%), and pefloxacin (20.4%). And it showed almost no cross-reactivity with other antibiotics or sulfonamides evaluated in this study. The competitive ELISA kit developed here could be used as a screening tool to detect and control illegal addition of CPFX in food products. This kit had been applied to milk detection and the recovery rates from samples spiked by CPFX were in a range of 63.02%–84.60%, with coefficients of variation of less than 12.2%.     

A study of the relationship between expression level of TRF1 protein and telomerase activity in human acute leukemia

INTRODUCTION Telomeres are distinctive DNA-protein struc-tures that cap the ends of linear chromosomes.It is very important to keep the chromosomes stabilization.Telomerase activity is closely linked to attainment of cellular immortality,a step in carcinogenesis,while lack of such activity contributes to cellular senes-cence.Telomerase is activated in more than85%ofmalignant tumors(Hiayma et al.,1997).Human te-lomeric repeat binding factor1(TRF1)is a telomere associated with proteins a…     

硅基材料的纳米结构设计和复合化及其在锂离子电池负极材料中的应用

锂离子电池因其具有能量密度大、电压高、比容量大、循环寿命长、环境友好等特点,被广泛应用于便携式电器和电动汽车等领域。锂离子电池的技术关键之一在于负极,开发高比容量、良好电化学性能的负极材料是锂离子电池研究的重要课题。近年来,硅基负极材料由于其极高的理论比容量,优异的大电流充放电性能等优点,受到了研究者们越来越多的关注。本文从硅基材料的纳米化结构设计及与其他材料的复合化两方面介绍了其作为锂离子电池负极材料的最新研究进展,并分析了目前存在的问题,指出了硅基材料作为锂离子电池负极材料的发展趋势。     

Changes of oxidative and lipoperoxidative metabolism in patients with acute viral myocarditis

Objective: To study the changes of oxidative metabolism and lipoperoxidative metabolism in patients with acute viral myocarditis (AVM), and to research pathological chain reactions of a series of free radicals, and oxidative damage and lipoperoxidative damage in AVM patients' bodies. Methods: A random paired control design was used to investigate 70 cases of AVM patients and 70 cases of healthy adult volunteers (HAV) on the basis of contents of nitric oxide (NO), lipoperoxide (LPO) in plasma, and LPO in red blood cell (RBC) as well as activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in RBC as revealed by spectrophotometric assays. Results: Compared with the average values (AV) of the above biochemical parameters (BP) in the HAV group, the AV of NO in plasma, and LPO in plasma and in RBC in the AVM group were significantly increased (P=0.0001), while the AV of SOD, CAT and GSH-Px in the AVM group were significantly decreased (P=0.0001). Conclusion: The findings in this study suggested that in the AVM patients bodies the metabolism of NO was disturbed, and the pathological chain reactions of a series of free radicals were so severely aggravated, as to cause the oxidative damage and lipoperoxidative damage in the AVM patients' bodies.     

HPAC及其研究进展

对高效亲和色谱技术八十年代以来的发展状况做了评述 ,介绍了高效亲和色普分离过程 ,评价了各种载体、配基、间隔臂及亲和色谱理论的特点 ,并对其未来进行了展望     

凝集素研究进展

凝集素包括微生物凝集素、植物凝集素、动物凝集素,它们在医药和农业科学领域中均有重要应用价值。不同来源凝集素的特点、分布及作用机理各不相同。凝集素的作用涉及细胞识别、微生物毒力决定子、植物抗虫、根瘤菌识别因子等诸多方面。     

发泡鞋底材料研究进展

综述了近年来发泡鞋底材料的研究现状和发展趋势,着重介绍了以聚氯乙烯(PVC)、乙烯-醋酸乙烯酯共聚物(EVA)、苯乙烯-丁二烯-苯乙烯嵌段共聚物(SBS)、聚氨酯(PU)为基体的几种典型发泡鞋底材料,并提出发泡鞋底材料应朝着功能化和环保化方向发展的建议。     

微铣削加工技术的发展现状

微铣削被认为是最柔性的加工方法之一,有着日益广阔的应用前景;介绍了微铣削加工技术的研究背景,并对其研究现状进行了评述,分析了微铣刀的特性和主要几何形状,并从切屑的形成、切削力和刀具磨损三个方面解析微铣削加工技术的加工机理及在研究与发展过程中存在的主要问题,讨论了微铣削技术的研究成果和发展前景。