Journal of chromatography最新文献_第6页

High-performance liquid chromatography of amino acids, peptides and proteins. CXXXII. Optimisation of operating parameters for protein purification with chromatographic columns. 氨基酸、多肽和蛋白质的高效液相色谱。CXXXII。色谱柱纯化蛋白质操作参数的优化。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87009-5

Q M Mao, I G Prince, M T Hearn

{"title":"High-performance liquid chromatography of amino acids, peptides and proteins. CXXXII. Optimisation of operating parameters for protein purification with chromatographic columns.","authors":"Q M Mao, I G Prince, M T Hearn","doi":"10.1016/s0021-9673(99)87009-5","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87009-5","url":null,"abstract":"In large-scale chromatography, process optimisation is one of the key elements for success. This paper presents a method for determining the optimum operating parameters for affinity and ion-exchange chromatographic columns when used for protein purification. Based on a mathematical model developed as part of our association investigations, computer programs have been developed to describe the dynamic relationships acting within the chromatographic system. Two basic operating parameters, the flow-rate and the effluent concentration at which the adsorption stage is terminated, can be optimised to give a maximum production rate. The sample loading volume and the processing time then can be determined. The effect of washing conditions on the production rate and the yield is also discussed. Examples are given for a specific system where the optimisation is based on the yield and the percentage utilisation of the column capacity. Contour plots are generated to aid the determination of the range of controlling parameters, and to guide further system design.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"81-9"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87009-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

High-performance liquid chromatography of amino acids, peptides and proteins. CXXX. Modified porous zirconia as sorbents in affinity chromatography. 氨基酸、多肽和蛋白质的高效液相色谱。CXXX。改性多孔氧化锆在亲和色谱中的吸附剂研究。

Journal of chromatography Pub Date : 1993-08-27 DOI: 10.1016/s0021-9673(99)87015-0

H J Wirth, M T Hearn

{"title":"High-performance liquid chromatography of amino acids, peptides and proteins. CXXX. Modified porous zirconia as sorbents in affinity chromatography.","authors":"H J Wirth, M T Hearn","doi":"10.1016/s0021-9673(99)87015-0","DOIUrl":"https://doi.org/10.1016/s0021-9673(99)87015-0","url":null,"abstract":"The utilisation of organosilanes to introduce active chemical groups onto zirconia surfaces, suitable for the subsequent immobilisation of proteins or other biomimetic ligands, is described. Two different types of porous zirconia-based particles with nominal pore diameters of 160 and 1000 A pore size were modified with two different affinity ligands. In the first case, methods to immobilise iminodiacetic acid-Cu(II) and its application in Cu(II) immobilised metal ion affinity chromatography (IMAC) were established. In the second series of experiments, concanavalin A was immobilised and the interaction of this lectin with the enzyme horseradish peroxidase examined. For both systems, adsorption isotherms were recorded as batch experiments. In each case, the experimental results could be fitted to langmuirean type adsorption isotherms, indicating that under the chosen conditions only one type of interaction is present, with nonspecific interactions with the support surface playing an insignificant role. These studies document the potential of surface modified zirconia particles for the immobilisation of chemical ligands or proteins for use in biospecific affinity chromatography and immobilised enzyme bioreactors.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"646 1","pages":"143-51"},"PeriodicalIF":0.0,"publicationDate":"1993-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/s0021-9673(99)87015-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19394028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 32

Characterization of the binding and chiral separation of D- and L-tryptophan on a high-performance immobilized human serum albumin column. D-色氨酸和l -色氨酸在高效固定化人血清白蛋白柱上结合和手性分离的表征。

Journal of chromatography Pub Date : 1993-08-20 DOI: 10.1016/0021-9673(93)83383-4

J Yang, D S Hage

{"title":"Characterization of the binding and chiral separation of D- and L-tryptophan on a high-performance immobilized human serum albumin column.","authors":"J Yang, D S Hage","doi":"10.1016/0021-9673(93)83383-4","DOIUrl":"https://doi.org/10.1016/0021-9673(93)83383-4","url":null,"abstract":"High-performance affinity chromatography was used to study the separation and binding of D- and L-tryptophan on an immobilized human serum albumin (HSA) column. Frontal analysis and zonal elution studies indicated that both D- and L-tryptophan were binding to single but distinct sites on HSA. L-Tryptophan bound to the indole site of HSA. D-Tryptophan had indirect interactions with the warfarin site of HSA but no interactions with the indole site. The association constants for the binding of D- and L-tryptophan at pH 7.4 and 25 degrees C were 0.4 x 10(4) and 2.7 x 10(4) M-1, respectively. The value of delta G for these sites ranged from -5.2 to -5.7 kcal/mol (1 cal = 4.184 J) and had a significant entropy component. The effects of varying the pH, phosphate concentration, temperature and polarity of the mobile phase on the binding of D- and L-tryptophan to HSA were examined. The role of sample size in determining peak shape and retention was also considered. From these data, general guidelines were developed for the separation of D- and L-tryptophan on immobilized HSA. Under optimized conditions the enantiomers were separated in less than 2 min with baseline resolution.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"645 2","pages":"241-50"},"PeriodicalIF":0.0,"publicationDate":"1993-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0021-9673(93)83383-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392206","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 104

Capillary zone electrophoresis of malto-oligosaccharides derivatized with 8-aminonaphthalene-1,3,6-trisulfonic acid. 8-氨基萘-1,3,6-三磺酸衍生麦芽糖的毛细管区带电泳。

Journal of chromatography Pub Date : 1993-08-20 DOI: 10.1016/0021-9673(93)83394-8

C Chiesa, C Horváth

{"title":"Capillary zone electrophoresis of malto-oligosaccharides derivatized with 8-aminonaphthalene-1,3,6-trisulfonic acid.","authors":"C Chiesa, C Horváth","doi":"10.1016/0021-9673(93)83394-8","DOIUrl":"https://doi.org/10.1016/0021-9673(93)83394-8","url":null,"abstract":"Malto-oligosaccharides were derivatized via their reducing end with 8-aminonaphthalene-1,3,6-trisulfonic acid by reductive amination, and the separation and electrophoretic migration behavior of the labelled sugars were investigated by capillary zone electrophoresis. Series of linear malto-oligosaccharides were found particularly suitable for both the study of the effect of the operating conditions on the separation and the investigation of the relationship between the electrophoretic mobility and the molecular size of the homologues. The electrophoretic mobility of the malto-oligosaccharide conjugates was found to be a linear function of the molecular mass to the negative two-thirds power. The sugar derivatives employed here carry three negative charges due to the presence of the dissociated sulfonic acid groups even at strongly acidic pH. Therefore, the analytes can migrate in the electric field without interference by electroendosmotic flow and/or wall adsorption in uncoated silica capillaries at low pH. As a result, the separation of these carbohydrate conjugates can be carried out under such conditions with high speed and efficiency in free solution, i.e., without an anticonvective medium such as a gel or a viscous polymer solution. Appropriate use of triethylammonium phosphate buffer, pH 2.5, as the background electrolyte improves not only the reproducibility, but also the efficiency and speed of the separation. The labelled sugars allow monitoring of the separation by UV detector or laser-induced fluorescence detector with concomitant enhancement of analytical sensitivity.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"645 2","pages":"337-52"},"PeriodicalIF":0.0,"publicationDate":"1993-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0021-9673(93)83394-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19392207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 153

Separation of isoforms of Serratia marcescens nuclease by capillary electrophoresis. 粘质沙雷氏菌核酸酶同工型的毛细管电泳分离。

Journal of chromatography Pub Date : 1993-08-20 DOI: 10.1016/0021-9673(93)83395-9

J Pedersen, M Pedersen, H Søeberg, K Biedermann

引用次数: 9

Binding characteristics of various neurochemicals to glassy carbon. 各种神经化学物质与玻碳的结合特性。

Journal of chromatography Pub Date : 1993-08-20 DOI: 10.1016/0021-9673(93)83381-2

Y Ikarashi, C L Blank, Y Maruyama

{"title":"Binding characteristics of various neurochemicals to glassy carbon.","authors":"Y Ikarashi, C L Blank, Y Maruyama","doi":"10.1016/0021-9673(93)83381-2","DOIUrl":"https://doi.org/10.1016/0021-9673(93)83381-2","url":null,"abstract":"The determination of acetylcholine and choline using liquid chromatography with electrochemical detection using the approach of Potter et al. [J. Neurochem., 41 (1983) 188] normally requires isolation of the desired species before an analysis of tissue samples can be undertaken due to coeluting interferences afforded by other neutrochemicals. We have recently shown that this problem can be overcome by the use of a glassy carbon precolumn to effectively trap the interfering species [Ikarashi et al., J. Chromatogr., 575 (1992) 29]. We now report on the nature and mechanism of this adsorption onto glassy carbon for norepinephrine, dopamine, serotonin, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid. For both the acidic and basic compounds which comprise this group, distinct Langmuir adsorption processes appear to be involved for both the neutral and ionic forms of the individual compounds. Using various data fitting approaches, we have attempted to derive appropriate adsorption constants for the two forms of each compound. Theoretical predictions employing these derived constants provided results which match reasonably well with the observed adsorption data in most cases.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"645 2","pages":"219-31"},"PeriodicalIF":0.0,"publicationDate":"1993-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0021-9673(93)83381-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19097966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 5

High-performance gel-permeation chromatographic analysis of protein aggregation. Application to bovine carbonic anhydrase. 蛋白质聚集的高效凝胶渗透色谱分析。应用于牛碳酸酐酶。

Journal of chromatography Pub Date : 1993-08-13 DOI: 10.1016/0021-9673(93)80623-g

C De Felice, K Hayakawa, T Watanabe, T Tanaka

引用次数: 8

Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography. 相对于膜层析，梯度反相高效液相色谱中蛋白质的区迁移和带展宽的特性。

Journal of chromatography Pub Date : 1993-08-13 DOI: 10.1016/0021-9673(93)80613-d

B G Belenkii, A M Podkladenko, O I Kurenbin, V G Mal'tsev, D G Nasledov, S A Trushin

{"title":"Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography.","authors":"B G Belenkii, A M Podkladenko, O I Kurenbin, V G Mal'tsev, D G Nasledov, S A Trushin","doi":"10.1016/0021-9673(93)80613-d","DOIUrl":"https://doi.org/10.1016/0021-9673(93)80613-d","url":null,"abstract":"The peculiarities of zone migration and band broadening in the reversed-phase gradient HPLC of proteins were investigated. In the isocratic mode a critical composition of the mobile phase was found at which all proteins regardless of their molecular mass migrate with equal velocity and have a capacity factor equal to the phase ratio (VP/V0), i.e., the same capacity factor as a marker of total accessible volume would have in steric exclusion chromatography. It is shown that steric exclusion conditions are never achieved in gradient HPLC. In the first (adsorption stage) of gradient elution where the separation takes place the velocity of a protein increases until it becomes equal to the velocity of the desorbing solvent front at a critical distance X0 from column entrance. Strong broadening is characteristic of this stage. In the second (critical) stage the protein travels the remaining distance (L-X0) with the velocity of the solvent. A definition of X0 is given allowing one very simple calculation of the minimum permissible column length as a function of gradient steepness, mobile phase velocity and protein adsorption parameter. When x = X0 the protein zone has the smallest dispersion. Making L < X0 is especially disadvantageous, as it leads to anomalous bandspreading. The theory of gradient HPLC was refined on this basis and the usefulness of this approach in high-performance membrane chromatography is demonstrated.","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"645 1","pages":"1-15"},"PeriodicalIF":0.0,"publicationDate":"1993-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0021-9673(93)80613-d","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19393526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 40

Quantitation of 5-methylcytosine by one-dimensional high-performance thin-layer chromatography. 一维高效薄层色谱法定量测定5-甲基胞嘧啶。

Journal of chromatography Pub Date : 1993-08-13 DOI: 10.1016/0021-9673(93)80635-l

S A Leonard, S C Wong, J W Nyce

引用次数: 13

Determination of total vitamin C in fruits by capillary zone electrophoresis. 毛细管区带电泳法测定水果中总维生素C含量。

Journal of chromatography Pub Date : 1993-08-13 DOI: 10.1016/0021-9673(93)80637-n

M Chiari, M Nesi, G Carrea, P G Righetti

引用次数: 51