共查询到19条相似文献,搜索用时 93 毫秒
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非病毒载体质粒DNA已被广泛应用于基因治疗和DNA疫苗,目前迫切需要开发其大规模制备和分离纯化方法。亲和色谱是一种高分辨率、高选择性的分离技术,在蛋白质、抗体、核酸等生物大分子的分离纯化方面显示了良好的应用前景。本文综述了亲和色谱技术在超螺旋质粒DNA分离纯化中的研究进展,总结了各种亲和色谱方法分离超螺旋质粒DNA的机理和优缺点,并展望了亲和纯化技术在质粒DNA生产和制备中的应用前景。 相似文献
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柱色谱分离纯化茉莉醛梁达文(广西玉林高等师范专科学校化学系玉林537000)茉莉醛是一种广泛用于洗涤剂、化妆品和食品的重要香料 ̄[1-4]。利用苯甲醛与庚醛在碱性条件下合成的茉莉醛,其主要副产物庚醛自缩物(2-正戊基-2-壬烯醛)对茉莉醛的香气质量影... 相似文献
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按照人金属硫蛋白-3(hMT-3)的基因序列,选用大肠杆菌偏爱的密码子合成了全长hMT-3基因,并将其插入大肠杆菌融合表达质粒pALEX的多克隆位点中,在谷胱甘肽-硫-转移酶(GST)下游与GST融合表达。通过异丙基-β-D-硫代半乳糖苷(IPTG)诱导在大肠杆菌表达菌株BL21(DE3)LysS中表达了与重金属离子镉结合的融合蛋白GST-Cd2+-hMT-3。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表明融合蛋白主要在超声上清液中。分别通过“先纯化、后酶切”和“亲和柱色谱原位酶切”两种方法纯化了Cd2+-hMT-3,比较了两种方法的纯化效率和得率,表明原位酶切法操作简便,较之“先纯化、后酶切”法减少了洗脱、透析、冻干等步骤,从而也减少了样品的损失,提高了样品的纯度和得率。从摇瓶培养菌液中纯化获得了结合有Cd2+的完整的人金属硫蛋白-3,得率为1.8%。氨基酸组成分析结果表明所获得的Cd2+-hMT-3不含芳香族氨基酸和组氨酸,符合金属硫蛋白的特征;直读电感耦合等离子体发射光谱分析其硫镉原子比为21∶(7.5±0.1),与理论值21∶7基本吻合。 相似文献
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兔金属硫蛋白的分离与纯化 总被引:1,自引:0,他引:1
兔皮下注射CdCl2诱导金属硫蛋白(Metallothionein,简称MT),取肝脏匀浆,用乙醇─氯仿混合液萃取后经SephdexG-50柱分离,得到混合型MT.进一步用纤维素弱碱性阴离子交换树脂进行拆分,并在SephadexG-25柱上脱盐,得到MT-1和MT-2两种亚型.经氨基酸组咸、凝胶电泳及HPLC柱层析等分析表明,所得的两种亚型具有高度的均一性. 相似文献
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活性炭吸附富集污染水中痕量硫双威及其气相色谱测定 总被引:2,自引:0,他引:2
用粒径300-400μm活性炭吸附富集环境水中痕量硫双威,气相色谱法直接测定,方法简便,快速,灵敏,准确,实用。研究了活性炭对硫双威的吸附性能及不同粒度活性炭对硫双威吸附效率的影响。进行了最佳预洗方法和最佳洗脱剂的选择。 相似文献
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《Analytical letters》2012,45(20):1647-1664
Abstract Three aluminas were investigated with five eluents containing various amounts of water. The first water layer adsorption can be described by the Freundlich's model whilst adsorption of the following water layers can be interpreted with the Brunauer, Emmet and Teller model. Energetic identity of the three aluminas was proved and eight active sites per nm2 were evidenced. Retention of solutes was interpreted with a logarithmic model in which the capacity factor logarithm varies linearly with the eluent water content logarithm. 相似文献
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Giulia Molino Marta Gandiglio Sonia Fiorilli Andrea Lanzini Davide Drago Davide Papurello 《Molecules (Basel, Switzerland)》2022,27(22)
Organic waste can be efficiently converted into energy using highly efficient energy systems, such as SOFCs coupled to the anaerobic digestion process. SOFC systems fed by biogenous fuels, such as biogas or syngas, suffer long-term stability due to trace compound impacts. It follows that, a mandatory gas cleaning section is needed to remove these pollutants at lower concentrations. This work investigates the adsorption mechanism for micro-contaminant removal through experimental results achieved using solid sorbents. Samples of different sorbent materials were analyzed in the laboratory to determine their performances in terms of sulfur (mainly hydrogen sulfide) and siloxanes (mainly D4-Octamethylcyclotetrasiloxane) adsorption capacities. The analysis shows that the chemical composition of the samples influences the adsorption of H2S (i.e., presence of calcium, iron, copper), while the effect of their textural properties mainly influences the adsorption of siloxane compounds, such as D4. A quantitative analysis was performed considering the influence of gas velocity on adsorption capacity. By increasing the biogas velocity (+45% and +89%), there was an indirect correlation with the H2S adsorption capacity (−27% and −44%). This identified an aspect related to the residence time required to be able to remove and retain the trace compound. The results obtained and summarized were used to develop a strategy for the removal of trace compounds in large-scale plants, e.g., for water purification. 相似文献
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疏水膜色谱法对生物大分子的快速纯化 总被引:2,自引:0,他引:2
首次采用自制的分别级合了辛基、丁基、苯基及聚乙二醇-4000的4种常用疏水基团的纤维素疏水膜色谱柱,以键合了辛基、苯基的SepharoseCL-4B凝胶柱为对照,考察了疏水膜色谱柱对牛血清白蛋白(BSA)的动态吸附容量及流速对吸附容量的影响。疏水膜色谱柱对蛋白及酶具有较好的疏水吸附及纯化作用,但吸附容量比相应的琼脂糖凝胶柱低得多。增大流速及降低蛋白溶液质量浓度对疏水膜色谱柱的吸附容量影响较小,这些性能使膜色谱柱非常适合于大体积低质量浓度蛋白溶液(如基团工程培养液)的分离纯化。 相似文献
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The orientation of antibodies, employed as capture molecules on biosensors, determines biorecognition efficiency and bioassay performance. In a previous publication we demonstrated for antibodies attached covalently to silicon that an increase in their surface amount Γ, evaluated with ellipsometry, induces changes in their orientation, which is traced directly using Time-of-Flight Secondary Ion Mass Spectroscopy combined with Principal Component Analysis. Here, we extend the above studies to antibodies adsorbed physically on a 3-aminopropyltriethoxysilane (APTES) monolayer. Antibodies physisorbed on APTES (0 ≤ Γ ≤ 3.5 mg/m2) reveal the Γ ranges for flat-on, side-on, and vertical orientation consistent with random molecular packing. The relation between orientation and Γ is juxtaposed for silicon functionalized with APTES, APTES modified with glutaraldehyde (APTES/GA) and N-hydroxysuccinimide-silane (NHS-silane). Antibody reorientation occurs at lower Γ values when physisorption (APTES) is involved rather than chemisorption (APTES/GA, NHS-silane). At high Γ values, comparable proportions of molecules adapting head-on and tail-on vertical alignment are concluded for APTES and the NHS-silane monolayer, and they are related to intermolecular dipole–dipole interactions. Intermolecular forces seem to be less decisive than covalent binding for antibodies on the APTES/GA surface, with dominant head-on orientation. Independently, the impact of glutaraldehyde activation of APTES on vertical orientation is confirmed by separate TOF-SIMS measurements. 相似文献
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用CNBr-activated Sepharose4B和微囊藻毒素-LR的单克隆抗体制备了免疫亲和层析柱,测得抗体偶联率在75.7%~94.1%之间。制得的免疫亲和层析柱最大柱容量在76~95ng之间,柱空白为0,回收率在90.8%~105.1%之间。柱子再生重复使用6次后,回收率不低于75%。建立了免疫亲和层析柱-高效液相色谱测定水样中的微囊藻毒素-LR的方法。该法检出限为5ng/L;线性定量范围为10~500ng/L。实验结果显示,免疫亲和层析柱特异性好,一次净化能除去绝大部分干扰物,净化效果明显优于现有的固相萃取柱。 相似文献
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Anion exchange chromatography is the most popular chromatographic method for plasmid separation.POROS R1 50 is a perfusio n chro mato graphic suppo rt w hich is a reversed phase matrix and is an alternative to co nventio nal o nes due to its mass transfer pro perties.The adso rptio n and elutio n o f the pIDKE2 plasmid o nto reversed phase POROS R1 50 w as studied.Langmuir iso therm mo del w as adjusted in o rder to get the max imum adso rptio n capacity and the disso ciatio n co nstant fo r POROS R1 50-plasmid DNA(pDNA) system.Breakthro ugh curves w ere o btained fo r vo lumetric flo w s betw een 0.69-3.33 mL/min,given dynamic capacity up to 2.3 times higher than tho se repo rted fo r io nic ex change matrix used during the purificatio n pro cess o f plasmids w ith similar size to that o f pIDKE2.The efficiency w as less than 45% fo r the flo w co nditio ns and initial co ncentratio n studied,w hich means that the suppo rt w ill no t be o perated under saturatio n circumstances. 相似文献