聚酰胺薄膜层析及其用于蛋白质化学分析

聚酰胺薄膜层析是1966年后发展起来的一种新层析技术。特别在用于分析氨基酸衍生物——DNS-氨基酸、DNP-氨基酸和PTH-氨基酸时,具有灵敏度高、分辨力强、速度快、操作方便等优点,超过了过去分析这类化合物使用的纸层析、纸电泳、硅胶薄板层析等方法。在蛋白质化学结构分析中,聚酰胺薄膜层析结     

食品安全免疫层析检测技术研发及应用进展

近年来,免疫层析检测技术在食品安全领域的发展迅速。本文简述了2015年国家食品安全法修订以来,我国食品安全快速检测产业相关的法律法规政策的制定和发展。回顾了免疫层析检测技术在我国的应用和发展历程,围绕免疫层析试纸条技术在食品安全检测领域的产品类型、检测方式、发展趋势等进行了分析和总结。评述指出,理论研究的进步和行业政策的扶持有力促进了我国免疫层析检测技术的壮大和产业化发展,但目前免疫层析试纸条产品自身的缺陷,产品评价方法缺乏以及政府主管部门监管体制的不完善等仍然是制约其健康发展的关键因素。文章最后展望了在大数据信息化时代,检测数据信息化平台的构建将进一步推动免疫层析检测技术在食品安全领域的发展。     

纤维素板微量氨基酸的分离及几种显色方法的比较

氨基酸分析是研究蛋白质化学的最基本的实验技术之一。现在已经普遍地使用离子交换层析、纸层析、纸电泳、薄层层析等方法分离、分析氨基酸。其中自动化的离子交换层析方法分析具有快速、微量和准确等特点,但需要昂贵的仪器设备,在一般实验室里不易办到。纸层析和薄层层析的方法,设备要求简单,操作方便,操作熟练时也可达到定量的要求。也可以用纤维素薄层层析的方法分离分析氨基酸,并已用于尿和血样品以及其他蛋白质中的氨基酸测定。经我们初步摸索,使用一种小型的纤维素板层析,并将茚三酮试剂直接加入展层溶剂中,蛋白质经酸水解后,可得到完全分开的普通     

红细胞生成刺激素(EPO)的纯化

 本文用中空纤维柱超滤浓缩尿,再经离子交换层析、聚焦层析、凝胶过滤和制备型聚丙烯酰胺凝胶电泳(PAGE)-层析等四步从15L再生障碍性贫血患者尿中获得约2mg EPO制品。比活性达10 300U/mg蛋白。该制品在分析型PAGE中呈一条区带。     

氨基酸定性分析技术的一些改进 Ⅱ.高压电泳高温层析双向纸上分离,小型滤纸层析和琼脂电泳

(1)本文以高压电泳和高溫层析分离氨基酸,可在10小时左右完成。甘氨酸与絲氨酸,組氨酸,精氨酸和賴氨酸的分离較双向层析为佳;但谷氨酸、門冬氨酸、谷氨酰胺与門冬酰胺尚需借第二向长时間层析才能分开。(2)以高溫层析溶剂系統作小型滤紙层析,可得高温层析图譜的縮影。小型滤紙层析的特点是样品和溶剂用量少,速度快,設备簡单,适合于大批样品同时并举之用。(3)利用pH8.8,离子強度0.025的巴比土緩冲液在琼脂板上进行DNP-谷氨酸,DNP-門冬氨酸,DNP-谷氨酰胺和DNP-門冬酰胺的电泳分离,效果很好,因此琼脂电泳法可作为末端分析的一种輔助技术,分辨上述4种DNP-氨基酸和驗証紙层析的結果。     

层析法纯化破伤风毒素的试验研究

为了获得高纯度的破伤风毒素,用疏水层析和离子交换层析纯化破伤风毒素。破伤风毒素培养滤液经Phenyl Sepharose疏水层析除去大部分杂质,再经DEAE Sephadex离子交换层析进一步纯化。经两步层析纯化后,毒素纯度达到2000Lf/mg PN以上,回收率为52%~73%。用此方法,连续纯化五批毒素,均获得高纯度的破伤风毒素。试验证明破伤风毒素经疏水层析和离子交换层析可得到有效纯化。     

菲利普斯公司新的分析目录册

<正> 新的1985～1986Philips 公司分析目录详述了该公司多种分析技术,其中包括有在上几个月中大批量新投产的信息。这本40页的发行本报道了原子吸收作用光分度计、扫描透射电镜、微量分析,发射光度计、气相层析、红外线分光光度计和液相层析的各种仪器。同时,也     

毕赤酵母表达重组人白细胞介素11的纯化与鉴定

报道了毕赤酵母表达人白细胞介素11的下游工艺研究,并对其产物进行了分析鉴定。所用工艺流程为离心收集上清、超滤浓缩脱盐、离子交换层析、疏水层析、凝胶过滤。所得产物经SDSPAGE电泳、RPHPLC分析、N端和C端序列分析、质谱、等电点分析和生物学活性分析,结果表明：产品纯度大于97%,结构和性质与E.coli融合表达的Neumega完全一致。     

层析液的配制

叶绿体中色素的提取和分离实验中用到层析液来分离色素,关于层析液的配制,高中《生物》课本２８８页有注释：教师在实验课前配制好层析液［层析液由２０份石油醚（６０～９０℃）、２份丙酮、１份苯混合而成］。有的老师在配制层析液时,按比例量取石油醚,将石油醚加热...     

三种展层剂层析效果的比较

植物色素的层析方法有多种。目前多采用纸层析、薄板层析和柱层析三种,其中以纸层析法最为常用。此法虽简便、经济、易行,但如展层剂选择不好,常会造成层析效果不佳。为提高色素的分离、提纯效果、增大各色素带间的距离,我们选用双重展层剂,分     

Components and antioxidant activity of the polysaccharide from Streptomyces virginia H03

A polysaccharide was isolated from the broth of cultured Streptomyces virginia H03 which was treated by ethanol deposition and savage method to remove the protein, and was purified using Sephadex G-150 column chromatography. The components of the polysaccharide were determined by gas chromatography. The purified polysaccharide was made up of mannose, glucose and galactose, in a 2:1:1 proportion. Its average apparent molecular weight was 3.76 x 10(4) Da which was determined by gel permeation chromatography. In addition, several antioxidant assays were adopted to investigate the antioxidant activity of the polysaccharide in vitro. The results indicated that the purified polysaccharide showed significant antioxidant activity against superoxide anion, hydrogen peroxide and 1,1-diphenyl-2-picrylhydrazyl radical, and lipid peroxidation as with standard antioxidants such as vitamin C. Furthermore, the polysaccharide had a better heat stability than vitamin C, which suggested that the polysaccharide might be a potent useful antioxidant.     

Isolation and purification of cell wall polysaccharide of Bacillus anthracis (Δ Sterne)

A polysaccharide fraction was isolated form sodium-dodecyl-sulfate (SDS) treated cell walls of Bacillus anthracis (delta Sterne) by hydrofluoric acid (HF) hydrolysis and ethanolic precipitation. The polysaccharide fraction was subsequently purified by several washings with absolute ethanol. Purity of the isolated polysaccharide was tested using the anthrone assay and amino acid analyzer. The molecular mass of the polysaccharide fraction as determined by gel filtration chromatography was about 12000 Da. Preliminary analyses of the polysaccharide was done using thin layer chromatography and amino acid analyzer, and results obtained from these analyses were further confirmed by gas liquid chromatography and 13C-NMR spectroscopy. Results showed that the polysaccharide moiety contained galactose, N-acetylglucosamine, and N-acetylmannosamine in an approximate molar ratio of 3:2:1. This moiety was devoid of muramic acid, alanine, diaminopimelic acid, glutamic acid, and lipid, thus indicating that the isolated polysaccharide was of pure quality.     

Isolation of a coaggregation-inhibiting cell wall polysaccharide from Streptococcus sanguis H1.

Coaggregation between Streptococcus sanguis H1 and Capnocytophaga ochracea ATCC 33596 cells is mediated by a carbohydrate receptor on the former and an adhesin on the latter. Two methods were used to release the carbohydrate receptor from the gram-positive streptococcus, autoclaving and mutanolysin treatment. The polysaccharide released from the streptococcal cell wall by either treatment was purified by ion-exchange chromatography; this polysaccharide inhibited coaggregation when preincubated with the gram-negative capnocytophaga partner. After hydrolysis of the polysaccharide by hydrofluoric acid (HF), the major oligosaccharide of the polysaccharide was purified by high-performance liquid chromatography. By analysis of the HF hydrolysis of the polysaccharide and the purified oligosaccharide, this major oligosaccharide appeared to be the repeating unit of the polysaccharide, with minor components resulting from internal hydrolysis of the major oligosaccharide. Gas chromatography results showed that the oligomer was a hexasaccharide, consisting of rhamnose, galactose, and glucose, in the ratio of 2:3:1, respectively. By weight, the purified hexasaccharide was a fourfold-more-potent inhibitor of coaggregation than the native polysaccharide. Resistance to hydrolysis by sulfuric acid alone and susceptibility to hydrolysis by HF suggested that oligosaccharide chains of the polysaccharide are linked by phosphodiester bonds. Studies with a coaggregation-defective mutant of S. sanguis H1 revealed that the cell walls of the mutant contained neither the polysaccharide nor the hexasaccharide repeating unit. The purification of both a polysaccharide and its constituent hexasaccharide repeating unit, which both inhibited coaggregation, and the absence of this polysaccharide or hexasaccharide on a coaggregation-defective mutant strongly suggest that the hexasaccharide derived from the polysaccharide functions as the receptor for the adhesin from C. ochracea ATCC 33596.     

阿魏蘑多糖理化性质及免疫活性研究*

以阿魏蘑Pleurotus ferulae Lanzi子实体和菌丝体为试验材料,采用水浸法提取阿魏蘑多糖,分别得到子实体粗多糖A和菌丝体粗多糖B。将A经Sevag法去蛋白、透析、CTAB络合、乙醇沉淀、NaCl溶液溶解、透析,得到多糖A1。紫外光谱分析鉴定多糖A1为均一组分。苯酚—硫酸法测得多糖A1糖含量为82.9%。凝胶渗透色谱法测得多糖A1数均分子量Mn=141088,重均分子量Mw=142897。气相色谱分析多糖A1单糖组成及其摩尔比为Xyl∶Gla∶Glc=1∶1.102∶2.899。巨噬细胞吞噬作用试验、迟发型变态反应试验、白细胞介素-2（IL-2）的诱生与检测试验测得粗多糖A、粗多糖B具有免疫活性。     

离子色谱在多糖疫苗及多糖蛋白结合疫苗质量控制中的应用

离子色谱具有方便、快速、灵敏度高、选择性好、重现性好、准确度高、可同时分析多种离子化合物等优点,其中的高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD)可直接分析糖类物质,可应用于多糖疫苗及多糖蛋白结合疫苗中多糖的分析。近年来,离子色谱应用在多糖及多糖蛋白结合疫苗的质量控制中,如测定疫苗中的多糖、游离糖、杂质多糖、对人有免疫原性的功能基团等的研究有许多报道,对有关内容进行了综述。     

斜生褐孔菌多糖组分的纯化及其生物活性研究

对斜生褐孔菌子实体和菌丝体的多糖含量进行了测定,二者多糖含量分别为2.63%和2.12%。运用DEAE Sephacel和Sephadex G-200柱层析技术从子实体中纯化出B1、B2、B31和B32四种多糖组分,分别占子实体多糖总量的50.20%、32.25%、4.48%和6.61%;从菌丝体中纯化出M1、M2和M3三种多糖组分,分别占菌丝体多糖总量的48.23%、31.85%和10.91%。生物活性试验结果显示:七种纯化多糖组分的抗氧化性由强到弱依次为B1>M1>M2>B2>M3>B32>B31;B2和M2具有在体外抑制人体白血病细胞株K-562和肝癌SMMC772l细胞株生长的作用;B31、B32和M3具有降低四氧嘧啶糖尿病小鼠血糖的作用,而且对健康小鼠的血糖值并无影响。     

淡紫拟青霉胞外多糖的分离、纯化及结构分析

淡紫拟青霉NH-PL-03菌株的胞外多糖粗提物对枯萎病病原菌-尖孢镰刀菌具有较好的抑制效果,文中对淡紫拟青霉胞外多糖进行了分离纯化和结构分析,以期为其构效关系研究奠定基础。采用乙醇沉淀法从淡紫拟青霉发酵液中提取粗多糖,经Sevage法脱蛋白后,过Superdex-G75凝胶层析柱分离得到胞外多糖EP-1。紫外分光法和Sephacryl S-200 HR凝胶层析柱检测EP-1为均一多糖,Sephacryl S-200柱层析测得EP-1的分子量为35.2 kDa,完全酸水解后纸层析检测EP-1的单糖组成中仅有葡萄糖,红外光谱、高碘酸氧化和Smith降解结果表明EP-1的化学结构是以β-(1,3)糖苷键连接而成的无分枝的葡聚糖。刚果红络合试验表明EP-1在稀的碱溶液中以3股螺旋构象存在。     

Purification of capsular polysaccharide from Neisseria meningitidis serogroup C by liquid chromatography

Neisseria meningitidis serogroup C capsular polysaccharide (MenCPS) is an important antigen against meningococcal infection. This paper describes a new purification methodology employing liquid chromatography that resulted in a polysaccharide showing the characteristics recommended by the World Health Organization for vaccine purposes. In this method, steps of the traditional procedure that yield low recovery and use toxic materials were modified. The present process consists in the following steps: (1) continuous flow centrifugation of the culture for removal of the cells; (2) supernatant concentration by tangential filtration (100 kDa cutoff); (3) addition of 0.5% DOC, heating to 55 degrees C during 30 min and tangential filtration (100 kDa cutoff); (4) anion exchange chromatography (Source 15Q) and (5) size exclusion chromatography (Sepharose CL-4B). The polysaccharide C fraction obtained in that way was dialyzed and freeze-dried. The structural identity of the polysaccharide was demonstrated by (1)H-NMR spectrometry.     

Studies on plant gums. Isolation and characterisation of the major polysaccharide from Neem (Azadirachta indica) gum

Azadirachta indica (neem) exudate gum was treated with pronase for 48 h followed by chromatography on TEAE-cellulose and the major polysaccharide was isolated. The polysaccharide covalently associated with remnant protein, was homogeneous as indicated by rechromatography on TEAE-cellulose, paper electrophoresis, gel chromatography under dissociating conditions on Bio-Gel P-l00 and P-300. The monosaccharide units, galactose, arabinose, glucuronic acid, fucose and glucosamine were present in a molar ratio of 86 ∶ 70 ∶ 30 ∶ 10 ∶ 1. Thirteen amino acids constituted the protein portion. The linkage between the polysaccharide and the protein was a glucosaminyl asparginyl bond. Limited hydrolysis showed that fucose and arabinose were at the non-reducing ends of the polysaccharide and galactose and glucuronic acid were in the central core.     

Isolation and characterization of a succinylated polysaccharide from the cell wall of Micrococcus agilis

A polysaccharide consisting of rhamnose, galactose, glucosamine and ester-linked succinic acid was extracted from the isolated cell walls of Micrococcus agilis by the hot water-phenol and 5% trichloroacetic acid (TCA) extraction methods. The hot water-phenol extractable polysaccharide accounted for 30% of the weight of the wall, with 23% by the TCA method. Phosphorus contents were less than 0.01% of the polysaccharide. Succinyl residues released by alkali treatment (0.1 N NaOH, 30 min, 37 degrees C) were identified by gas-liquid chromatography, and accounted for 6.3% and 5.1% of the polysaccharide purified from the hot water-phenol and TCA extracts, respectively. The polysaccharide was not bound when chromatography on Concanavalin A-Sepharose 4B (Con A/Sepharose 4B) columns was performed and it could thus be separated from any residual membrane lipomannan. The purified polysaccharide behaved as a negatively-charged polymer on electrophoresis in 1% agarose (at pH 8.6). A strong cross-reaction, unaffected by removal of the succinyl groups, was observed with type XXIII pneumococcal polysaccharide antiserum indicating the presence of L-rhamnose, linked through non-reducing, lateral end groups.