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1.
Facile analysis and purification of deblocked N-terminal pyroglutamyl peptides with a strong cation-exchange sulfoethyl aspartamide column 总被引:1,自引:0,他引:1
D L Crimmins D W McCourt B D Schwartz 《Biochemical and biophysical research communications》1988,156(2):910-916
A high-performance strong cation-exchange Sulfoethyl Aspartamide column was used to analyze and purify five N-terminal pyroglutamyl peptides after treatment with Pyroglutamate Aminopeptidase. The resulting deblocked N-1 peptides possess an increased positive charge and are therefore retained to a greater extent by the column. Salt gradient elution in a pH 3 mobile phase was then used to recover the desired peptides and the purified deblocked peptides were directly subjected to N-terminal sequence analysis. The same digests were also chromatographed on a C18 reversed-phase column using standard trifluoroacetic acid-acetonitrile gradient elution. The elution order for the parent peptide and the N-1 peptide on the reversed-phase column was reversed from that on the Sulfoethyl Aspartamide column and the resolution of the two peptides obtained on the reversed-phase column was less than that observed on the cation-exchange column. In addition, the Sulfoethyl Aspartamide column was shown to be useful to monitor the extent of N-terminal glutamine cyclization formed during peptide purification and storage. 相似文献
2.
Column chromatographic separation of acidic and neutral and basic metabolites of tyrosine and dihydroxyphenylalanine 总被引:1,自引:0,他引:1
Liquid column chromatographic techniques for separation of tyrosine; dihydroxyphenylalanine; and acidic, neutral, and basic metabolites of tyrosine and catecholamines are described. These techniques permitted simultaneous separation of a wide spectrum of acidic and neutral metabolites of tyrosine and dihydroxyphenylalanine on one column. Rapid and complete separation of basic metabolites was carried out on a second column. 相似文献
3.
Siu SC Baldascini H Hearle DC Hoare M Titchener-Hooker NJ 《Bioprocess and biosystems engineering》2006,28(6):405-414
This study examined the impact of fouling with yeast homogenate on capacity and breakthrough performance of an ion exchange packed bed column. Column performance was assessed by analysis of breakthrough curves obtained with BSA as a test protein. The overall impact of fouling on breakthrough performance depended heavily on the level of clarification of the feed stream. Challenging the column with particulate-free homogenate caused no change in column performance. Loading successive small volumes of poorly clarified homogenate, interspersed with frequent column salt washes, did not alter significantly the column capacity. By contrast, when the column was challenged with an equivalent cumulative volume of poorly clarified homogenate, dynamic binding capacity decreased significantly and changes in breakthrough curves suggested increased intraparticle and external mass transfer limitations. These changes were ascribed to deposition of solid particulates in void spaces in the bed and colloidal contaminants in the bead pores. 相似文献
4.
Yang G Feng S Liu H Yin J Zhang L Cai L 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2007,854(1-2):85-90
A weak ion exchange monolithic column prepared by modifying the GMA-MAA-EDMA (glycidyl methacrylate-methacrylic acid-ethylene glycol dimethacrylate) monoliths with ethylenediamine was applied to remove matrix compounds in biological fluid. Using this monolithic column, on-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma samples had been investigated. Chromatography was performed by reversed-phase HPLC on a C(18) column with ultraviolet detection at 225 nm. Results showed that the ion exchange monolithic column could be used for deproteinization and retaining oxacillin and cloxacillin in human urine and plasma, which provided a simple and fast method for assaying drugs in human urine and plasma. 相似文献
5.
Function and attachment of the stem in Isocrinidae and Pentacrinitidae: review and interpretation 总被引:1,自引:0,他引:1
H. WIENBERG RASMUSSEN 《Lethaia: An International Journal of Palaeontology and Stratigraphy》1977,10(1):51-57
A review is given of observations and interpretations of the attachment in Isocrinida. It is concluded that all adult Isocrinida discard distal parts of their column and live attached by distal cirri to the bottom or to objects lying on the bottom. From time to time the column may break below a nodal and the isocrinid drift along the bottom until it regains attachment with the cirri. In undisturbed environments a considerable length of the tapering distal part of the column may be maintained, and several specimens may be connected by the entangled distal parts of the columns lying on the bottom. Specimens of Seirocrinus subangularis attached to tree trunks in the Lower Jurassic Posidonia shale of Holzmaden, Germany, have been interpreted as pseudoplanktic. These are all adult specimens with distal parts of their column discarded before they attached themselves to the trunks. Furthermore, since adult specimens cannot swim to the surface, they must have attached themselves to sunken trunks lying on the bottom. 相似文献
6.
研究了用固相萃取技术结合氢火焰气相色谱法测定水和废水中四种氯苯类有机污染物。通过实验比较Sdex C18,Sep-Park Vac Silica,Bond Elut CARBON,Bond Elut SI和Bond Elut PLEXA五种SPE小柱对四种氯苯类的萃取效率,发现Dionex的Sdex C18柱有很好的回收率。系统研究了最佳萃取条件,甲醇洗脱体积为4.0 mL,且洗脱液不可通过氮吹浓缩;四种氯苯类化合物除氯苯外穿透体积都在1.0 L以上,而氯苯的穿透体积为300 mL,表明Sdex C18柱对二氯苯和三氯苯有很强的吸附性。在最佳萃取和测定条件下,方法线性范围为20.0~400μg/L,检出限为0.383~0.635μg/L,完全满足日常环境监测分析要求。 相似文献
7.
5-Hydroxymethylfurfural (HMF) and furfural could be separated by the Aminex HPX-87H column chromatography, however, the separation and quantification of acetic acid and levulinic acid in biomass hydrolysate have been difficult with this method. In present study, the HPLC separation of acetic acid and levulinic acid on Aminex HPX-87H column has been investigated by varying column temperature, flow rate, and sulfuric acid content in the mobile phase.The column temperature was found critical in resolving acetic acid and levulinic acid. The resolution for two acids increased dramatically from 0.42 to 1.86 when the column temperature was lowered from 60 to 30 °C. So did the capacity factors for levulinic acid that was increased from 1.20 to 1.44 as the column temperature dropped. The optimum column temperature for the separation was found at 45 °C. Variation in flow rate and sulfuric acid concentration improved not as much as the column temperature did. 相似文献
8.
Volume 61, no. 4, p. 1624, column 2, lines 38-41: The sentence should read "For example, at position 21, the G nucleotide (Fig. 1) was present in all the ISR B. thuringiensis subspecies except for B. thuringiensis subsp. tenebrionis (Te4), which contained an A." Page 1624, column 2, line 45: "Position 62" should read "position 11." Page 1624, column 2, line 47: "Position 90" should read "position 39." Page 1624, column 2, line 49: "Position 83" should read "position 32." Page 1625, column 1, line 3: "Position 83" should read "position 32." Page 1626, column 1, line 1: "Positions 62, 90, and 165, and one deletion at position 83" should read "positions 11, 39, and 114, and one deletion at position 32." [This corrects the article on p. 1623 in vol. 61.]. 相似文献
9.
De Cremer K Cornelis R Strijckmans K Dams R Lameire N Vanholder R 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2002,775(2):143-152
The behaviour of free [48V]vanadate and [48V]vanadium-transferrin complex was investigated on five different anion-exchange columns (Mono Q 5/5 HR, Hitrap Q HP, Sepharose Q FF, Sepharose DEAE FF and Hitrap Q XL). The recovery of both V-compounds was quantitative. The peak shape and retention time of vanadate varied according to the type of column. The vanadium-transferrin complex also showed different elution patterns depending on the type of column. Especially in case of the Sepharose Q FF, Mono Q 5/5 HR and Hitrap Q XL columns the vanadium-transferrin binding was degraded during elution on the column. The results clearly prove that care should be taken as to the choice of column for speciation purposes of vanadium compounds in order to prevent various artefacts showing up in the chromatograms. A Hitrap Q HP column was used to fractionate different vanadium compounds in rat serum. 相似文献
10.
Kazoka H 《Journal of biochemical and biophysical methods》2007,70(1):15-21
The chromatographic behavior of some purines and pyrimidines on a monolithic Chromolith Performance Si column under normal-phase high-performance liquid chromatography mode has been studied. Column pressure, column efficiency and selectivity of Chromolith Performance Si column were compared to those of conventional spherical 5 microm silica packed columns Econosphere Silica and Zorbax Rx-SIL. The investigation has shown that application of Chromolith Performance Si column for analysis of polar solutes can reduce the separation time without sacrificing column efficiency and selectivity. Improvement of the monolithic silica column efficiency for polar solutes is observed when ternary mobile phases (mixtures of hexane-isopropanol with ethylene glycol, water or acetonitrile) are applied. 相似文献
11.
Redox indicators were employed to monitor redox status in a bioaugmented, sediment-packed column during the dechlorination of tetrachloroethene (PCE) to ethene (ETH). The speciation of the indicators thionine and cresyl violet, immobilized on transparent films, was spectrometrically monitored with a flow sensor based on circulating the column solution through a specially constructed flow cell placed in a conventional spectrometer. A fiber optic redox probe based on immobilized azure C was constructed. A 75-cm column with 4 sets of ports along the column axis at regular intervals was constructed and packed with aquifer material. These ports enabled sampling to determine concentrations of chlorinated ethene species and allowed for in situ or non-invasive monitoring of redox conditions with negligible O 2 contamination. The flow sensor and fiber optic sensor showed similar responses to redox conditions in the column. After 60 days, complete conversion of PCE to ETH occurred by the end of the column and the redox level indicated by the indicators was consistent throughout the column. Significant formation of vinyl chloride or ETH was observed only after significant reduction of cresyl violet. 相似文献
12.
IgGPH_7经木瓜蛋白酶消解产生Fab和Fc,在FPLC仪上,分别用MonoQ,Superose_(12),MonoS柱提纯Fab。提纯的Fab经还原和烷基化后,再用琥珀酸酐修饰,然后用Mono Q柱分离得到轻链和重链。最后通过SDS凝胶电泳和N端顺序测定,以鉴定它的纯度和轻、重链。 相似文献
13.
A method is described for the purification of the dye azure B in quantities sufficient for biological staining experiments on a larger scale. The method is based on the use of column chromatography. Two columns are employed. In column A with silica gel as adsorbent the azure B fraction is isolated from a suitable substrate ('technical' azure B gained by a modification of Bernthsen's synthesis of methylene blue, or polychrome methylene blue) using an acetate-formate mixture as eluent. In column B, on an Amberlite polymeric adsorbent (XAD-2) the acetate-formate anions are exchanged in chloride. Regeneration of both columns is possible: KMnO4, Na2S2O4 and water are run through column A; 5% NaOH, methanol and water through column B. Purification of azure B on economic terms is thus attained. The opinion is expressed that this method is also applicable to the purification of other cationic dyes. 相似文献
14.
《仿生工程学报(英文版)》2008
Based on the configuration principles of biological skeletons and sandwich stems, a machine tool column with stiffening ribs inside was designed using structural bionic method. After the lightening effect was verified by finite element simulation, scale-down models of a conventional column and a bionic column were fabricated and tested. Results indicate that the bionic column can reduce the maximum static displacement by 45.9% with 6.13% mass reduction and its dynamic performances is also better with increases in the first two natural frequencies. The structural bionic design is effective in improving the static and dynamic structural performances of high speed machine tools. 相似文献
15.
A method is described for the purification of the dye azure B in quantities sufficient for biological staining experiments on a larger scale. The method is based on the use of column chromatography. Two columns are employed. In column A with silica gel as adsorbent the azure B fraction is isolated from a suitable substrate ('technical' azure B gained by a modification of Bernthsen's synthesis of methylene blue, or plychrome methylene blue) using an acetate-formate mixture as eluent. In column B, on an Amberlite polyineric adsorbent (XAD-2) the acetate-formate anions are exchanged for chloride. Regeneration of both columns is possible: KMnO4, Na2S2O4 and water are run through column A, 5% NaOH, methanol and water through column B. Purification of azure B on economic terms is thus attained. The opinion is expressed that this method is also applicable to the purification of other cationic dyes. 相似文献
16.
为探讨有尾目脊椎的划分,本文以小鲵科的山溪鲵(Batrachuperus pinchonii)为例,运用透明骨骼双色法对其脊柱的形态特征进行了观察,并对各部分椎骨特征进行详细描述和绘图.结果显示,山溪鲵的脊椎根据椎骨是否具前关节突、横突、肋骨、肋软骨和脉弓等形态特征可分5部分;同时结合小鲵科其他20种94号标本和蝾螈科6种27号标本的脊柱特征及文献资料,讨论了有尾目脊椎的划分,认为将有尾目脊柱划分为5部分(颈椎、躯椎、荐椎、尾荐椎和尾椎)的观点较将其划分为4部分(颈椎、躯椎、荐椎和尾椎)的观点更合理. 相似文献
17.
《Journal of biochemical and biophysical methods》2008,70(1):15-21
The chromatographic behavior of some purines and pyrimidines on a monolithic Chromolith Performance Si column under normal-phase high-performance liquid chromatography mode has been studied. Column pressure, column efficiency and selectivity of Chromolith Performance Si column were compared to those of conventional spherical 5 μm silica packed columns Econosphere Silica and Zorbax Rx-SIL. The investigation has shown that application of Chromolith Performance Si column for analysis of polar solutes can reduce the separation time without sacrificing column efficiency and selectivity. Improvement of the monolithic silica column efficiency for polar solutes is observed when ternary mobile phases (mixtures of hexane–isopropanol with ethylene glycol, water or acetonitrile) are applied. 相似文献
18.
Wang Y Xu Y Zhang X Wang E Dong Y 《International journal of biological macromolecules》2011,49(3):428-432
The development of a chitosan-supported immunoaffinity chromatography (IAC) column and its application to the selective extraction of methandrostenolone (MA) from food and feed samples were described in this paper. Using hybridoma technique, a monoclonal antibody (mAb) against MA was produced. The IAC column was prepared by coupling the produced antibody with crosslinked chitosan. Scanning electron microscopy and IR spectroscopy was used to characterize the chitosan crosslinking and antibody coupling. 2% and 90% methanol were respectively selected as loading and eluting solution by optimization. The maximum capacity of the column for MA was 1790 ng/mL gel. The extraction recoveries of the column for MA at three different spiked concentrations ranged from 83.7 to 98.5%. After 2 cycles of usage, the column capacity and extraction recovery still remained 84.6% and 80.5%. To further verify the effect of matrix on the IAC cleanup, MA-fortified food and feed samples were extracted using the prepared IAC column, and MA recovery rates were found to be 86.2% and 70.4%, respectively. 相似文献
19.
Purification and chemical characterization of human hexosaminidases A and B. 总被引:11,自引:9,他引:2
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N-Acetyl-beta-hexosaminidases A and B were purified to homogeneity from human placenta. In the initial step of purification, the enzymes were adsorbed on concanavalin A-Sepharose 4B and eluted from the column with alpha-methyl D-mannosides. Subsequent purification steps included DEAE-cellulose column chromatography, QAE-Sephadex [diethyl-(2-hydroxypropyl)aminoethyl-Sephadex] column chromatography, Sephadex G-200 gel filtration and preparative disc polyacrylamide-gel electrophoresis, followed by another QAE-Sephadex chromatography for the hexosaminidase A preparation, and DEAE-cellulose column chromatography, calcium phosphate gel chromatography, Sephadex G-200 gel filtration, QAE-Sephadex chromatography and CM-cellulose chromatography for the hexosaminidase B preparation. The purified preparations, particularly hexosaminidase A, had significantly higher specific enzyme activities than previously reported. The preparations moved on polyacrylamide-gel electrophoresis as single protein bands, which also stained for enzyme activity. Sedimentation-equilibrium centrifugation indicated homogenous dispersion of the enzymes, and the molecular weight was estimated as about 110000 for both enzymes. Complete amino acid and carbohydrate compositions of the two isoenzymes were determined, and, in contrast with previous suggestions, no sialic acid was found in the enzymes. 相似文献
20.
《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1991,570(1):99-107
A fully automated technique for high-performance liquid chromatographic analysis of whole blood and plasma is described. Samples are automatically injected into a dialyser where proteins and blood cells are removed. The dialysates are concentrated on a small column prior to analysis. This technique is used for the determination of oxytetracycline in whole blood and plasma. After dialysis oxytetracycline and the internal standard, tetracycline, are retained on a polystyrene enrichment column and subsequently separated on a polystyrene analytical column by ion-pair chromatography. Using ultraviolet detection 50 ng/ml can be detected. Validation showed good within-day and between-day accuracy and precision. Different oxytetracycline concentrations were found in plasma and whole blood. This difference varied between the species. 相似文献