Beyond labels: A review of the application of quantum dots as integrated components of assays, bioprobes, and biosensors utilizing optical transduction

A comprehensive review of the development of assays, bioprobes, and biosensors using quantum dots (QDs) as integrated components is presented. In contrast to a QD that is selectively introduced as a label, an integrated QD is one that is present in a system throughout a bioanalysis, and simultaneously has a role in transduction and as a scaffold for biorecognition. Through a diverse array of coatings and bioconjugation strategies, it is possible to use QDs as a scaffold for biorecognition events. The modulation of QD luminescence provides the opportunity for the transduction of these events via fluorescence resonance energy transfer (FRET), bioluminescence resonance energy transfer (BRET), charge transfer quenching, and electrochemiluminescence (ECL). An overview of the basic concepts and principles underlying the use of QDs with each of these transduction methods is provided, along with many examples of their application in biological sensing. The latter include: the detection of small molecules using enzyme-linked methods, or using aptamers as affinity probes; the detection of proteins via immunoassays or aptamers; nucleic acid hybridization assays; and assays for protease or nuclease activity. Strategies for multiplexed detection are highlighted among these examples. Although the majority of developments to date have been in vitro, QD-based methods for ex vivo biological sensing are emerging. Some special attention is given to the development of solid-phase assays, which offer certain advantages over their solution-phase counterparts.     

封闭群东方田鼠染色体制备中ConA的应用技术研究

人类染色体的制备常常采用外周血培养技术。但啮齿类动物的外周血培养以获得制备染色体的中期淋巴细胞比较困难 ,其原因不明。因此 ,啮齿类动物的染色体制备较多通过秋水仙素的作用 ,控制正在分裂的骨髓细胞 ,以获得中期分裂相的骨髓细胞 ,通过骨髓细胞来制备染色体。但发现用这种方法制备东方田鼠的染色体时 ,骨髓中处于中期分裂状况的中期细胞较少 ,不便于分析染色体。ConA是一种常用的有丝分裂刺激素 ,能有效地诱导T细胞增殖和分裂 ,是体外刺激小鼠等啮齿类动物T细胞分裂的必不可少的有丝分裂原。这种有丝分裂原在体内 ,特别是在东方田…     

金属卟啉与刀豆蛋白A相互作用的室温磷光研究

实验在脱氧胆酸钠盐(NaDC)介导下,采用室温磷光法研究阴离子meso-四-4-(羧基苯基)钯卟啉(Pd-TCPP)与刀豆蛋白A(ConA)之间的相互作用,实验表明Pd-TCPP与ConA之间能形成复合物,结合常数为3.33×10~4L·mol~(-1).这一结果可用于利用Pd-TCPP作为室温磷光探针对ConA进行测定.同时,紫外可见光谱和共振光散射先谱也同样证明Pd-TCPP与ConA之间存在相互作用.     

核桃楸青果皮(GPJM)水提物诱生小鼠肿瘤坏死因子的实验研究

核桃楸青果皮（GPJM）经沸水煮提得水提物，应用MTT法分析，GPJM水提物在体外可协同亚剂量ConA(2.5μg/mL)诱导小鼠脾细胞合成TNF；在体内也有诱生内源性TNF的作用。这表明核桃楸青果皮水提物（GPJM）具有一定的免疫调节作用。     

基于拟糖蛋白的电化学阻抗生物传感器用于糖与蛋白质相互作用的研究

以D-甘露糖和伴刀豆球蛋白(ConA)的相互作用为研究对象,利用美拉德反应将D-甘露糖共价结合到负载蛋白牛血清蛋白(BSA)的表面形成拟糖蛋白,然后将拟糖蛋白固定到玻碳电极表面,以拟糖蛋白表面的D-甘露糖为分子识别物质,构建了检测伴刀豆球蛋白(ConA)的电化学阻抗传感器。拟糖蛋白制备过程简单,D-甘露糖负载量大,在空间中提供多个结合位点,因此能与ConA形成多价复合物,提高了传感器的灵敏度。该传感器的响应值与ConA浓度的对数在5.0×10-11~5.0×10-9mol/L之间呈良好的线性关系,检出限为1.7×10-11mol/L,D-甘露糖和ConA之间的结合常数为2.6×106L/mol。该方法简单,可适用于不同糖和蛋白质相互作用的研究,为构建高灵敏度的电化学阻抗传感器提供了新思路。     

枸骨叶不同溶媒萃取物对小鼠体外T淋巴细胞活化增殖的影响

目的:探讨枸骨叶5种溶媒萃取物对T淋巴细胞增殖、活化的影响。方法:利用MTT法检测枸骨叶对体外ConA刺激T淋巴细胞增殖反应及使用流式细胞仪测定CD3阳性淋巴细胞的CD69分子表达的影响。结果:枸骨叶的醇提物I、乙酸乙酯IⅠ、正丁醇萃取物IⅡ具有明显抑制Co-nA刺激T淋巴细胞增殖反应的作用。枸骨叶醇提物I与乙酸乙酯萃取物IⅠ能明显抑制ConA刺激T淋巴细胞CD69分子的表达。枸骨叶2种水提物无免疫抑制作用。结论:枸骨叶脂溶性萃取物具有较强抑制T淋巴细胞活化、增殖的化学成分。     

伴刀豆球蛋白ConA体外诱导口腔鳞癌细胞Tca8113凋亡的机制初探

分别用0 μg/mL到100 μg/mL的伴刀豆球蛋白ConA加入人口腔鳞癌Tca8113细胞,采用MTT法检测ConA对Tca8113细胞的增殖抑制作用,并利用形态学观察、乳酸脱氢酶活力测定法(LDH)和流式细胞仪测定细胞凋亡,最后进行了caspase活性检测.加入25μg/mL的ConA培养24 h后,细胞的生长抑...     

N-glycosylation proteome of endoplasmic reticulum in mouse liver by ConA affinity chromatography coupled with LTQ-FT mass spectrometry

Glycosylation is the most versatile and one of the most significant protein post-translational modifications. It is generally classified into three categories according to the amino acid to which the glycan is attached: N-glycosylation, O-glycosylation and C-glycosylation. Synthesis of N-glycoproteins occurs in the rough endoplasmic reticulum (rER), and all N-glycoproteins synthesized in rER have uniform glycan endings with mannose (Man) and glucose (Glc). A systematic strategy was developed to comprehensiv...     

ConA或PHA对鮎(Silurus asotus)皮肤和鳃粘液细胞密度的影响

用阿利新蓝-过碘酸雪夫氏试剂染色法(AB-PAS),对鲇(Silurus asotus)皮肤和鳃粘液细胞进行分类,研究ConA或PHA浸泡和注射刺激对鲇皮肤和鳃粘液细胞密度变化的影响.结果表明:(1)除鳃丝软骨Ⅴ型粘液细胞密度经浸泡刺激无显著增加外,无论采取ConA或PHA浸泡刺激还是注射刺激,皮肤中的Ⅰ型,Ⅳ型粘液细胞和鳃中的Ⅱ型和Ⅲ型粘液细胞密度都有显著增加.(2)经ConA注射或浸泡刺激后,皮肤中粘液细胞密度分别增加了64.2%和145%,鳃丝中粘液细胞密度分别提高了173%和220%;经PHA注射或浸泡刺激后,皮肤粘液细胞密度分别增加了67.8%和142%,鳃丝粘液细胞密度分别增加了108%和196%;经ConA或者PHA注射刺激,鳃丝软骨的粘液细胞密度都有显著增加,分别增加了68.8%和13.2%,浸泡刺激对粘液细胞密度都没有显著影响(3)ConA刺激较PHA刺激效果明显,浸泡刺激较注射刺激效果明显.