Marı́a Iranzo, Antonio Marcilla, Marı́a Victoria Elorza, Salvador Mormeneo, Rafael Sentandreu
{"title":"亲和色谱法制备抗真菌细胞壁蛋白和抗甘露聚糖抗血清","authors":"Marı́a Iranzo, Antonio Marcilla, Marı́a Victoria Elorza, Salvador Mormeneo, Rafael Sentandreu","doi":"10.1006/emyc.1994.1017","DOIUrl":null,"url":null,"abstract":"<div><p>Iranzo, M., Marcilla, A., Elorza, M. V., Mormeneo, S., and Sentandreu, R. 1994. Preparation of anti-protein and anti-mannan antisera against fungal cell wall by affinity chromatography. <em>Experimental Mycology</em> 18, 159-167. A novel and easy chromatographic method has been developed for the isolation of anti-protein and anti-mannan antisera from a population of polyclonal antibodies obtained against <em>Candida albicans</em> and <em>Yarrowia lipolytica</em> cell wall mannoproteins. The technique is based on the immobilization of mannan (to be used as immunoadsorbent) by Affi-Prep H<sub>z</sub> resin after the oxidation of neighboring hydroxyl groups of the polysaccharide with sodium periodate. For <em>Y. lipolytica</em> polyclonal antiserum, a single chromatographic step using the homologous mannan was sufficient to obtain an antiprotein antibody preparation free of antimannan antibodies. For <em>C. albicans</em>, three chromatographic processes using homologous and heterologous mannan were needed to obtain a satisfactory antiprotein antiserum. The potential application of the anti-protein antiserum obtained has been demonstrated by indirect immunofluorescence assays of whole cells and electrophoretic analysis of wall proteins in <em>C. albicans</em> and <em>Saccharomyces cerevisiae</em> .</p></div>","PeriodicalId":12110,"journal":{"name":"Experimental Mycology","volume":"18 2","pages":"Pages 159-167"},"PeriodicalIF":0.0000,"publicationDate":"1994-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1006/emyc.1994.1017","citationCount":"2","resultStr":"{\"title\":\"Preparation of Anti-protein and Anti-mannan Antisera against Fungal Cell Wall by Affinity Chromatography\",\"authors\":\"Marı́a Iranzo, Antonio Marcilla, Marı́a Victoria Elorza, Salvador Mormeneo, Rafael Sentandreu\",\"doi\":\"10.1006/emyc.1994.1017\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Iranzo, M., Marcilla, A., Elorza, M. V., Mormeneo, S., and Sentandreu, R. 1994. Preparation of anti-protein and anti-mannan antisera against fungal cell wall by affinity chromatography. <em>Experimental Mycology</em> 18, 159-167. A novel and easy chromatographic method has been developed for the isolation of anti-protein and anti-mannan antisera from a population of polyclonal antibodies obtained against <em>Candida albicans</em> and <em>Yarrowia lipolytica</em> cell wall mannoproteins. The technique is based on the immobilization of mannan (to be used as immunoadsorbent) by Affi-Prep H<sub>z</sub> resin after the oxidation of neighboring hydroxyl groups of the polysaccharide with sodium periodate. For <em>Y. lipolytica</em> polyclonal antiserum, a single chromatographic step using the homologous mannan was sufficient to obtain an antiprotein antibody preparation free of antimannan antibodies. For <em>C. albicans</em>, three chromatographic processes using homologous and heterologous mannan were needed to obtain a satisfactory antiprotein antiserum. The potential application of the anti-protein antiserum obtained has been demonstrated by indirect immunofluorescence assays of whole cells and electrophoretic analysis of wall proteins in <em>C. albicans</em> and <em>Saccharomyces cerevisiae</em> .</p></div>\",\"PeriodicalId\":12110,\"journal\":{\"name\":\"Experimental Mycology\",\"volume\":\"18 2\",\"pages\":\"Pages 159-167\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1994-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1006/emyc.1994.1017\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Experimental Mycology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0147597584710176\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental Mycology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0147597584710176","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 2
摘要
Iranzo, M., Marcilla, A., Elorza, M. V., Mormeneo, S.和Sentandreu, R. 1994。亲和层析法制备抗真菌细胞壁蛋白和抗甘露聚糖血清。真菌学学报,18(2):559 - 567。建立了一种新的简便的色谱分离方法,用于从抗白色念珠菌和多脂耶氏菌细胞壁甘露糖蛋白的多克隆抗体中分离抗蛋白和抗甘露糖抗血清。该技术是在甘露聚糖的邻羟基被高碘酸钠氧化后,用Affi-Prep Hz树脂固定甘露聚糖(用作免疫吸附剂)。对于脂肪瘤多克隆抗血清,使用同源甘露聚糖的单层析步骤足以获得不含抗甘露聚糖抗体的抗蛋白抗体制剂。对于白色念珠菌,采用同源和异源甘露聚糖进行三次色谱处理,获得满意的抗蛋白抗血清。通过对白色念珠菌和酿酒酵母的全细胞间接免疫荧光分析和壁蛋白电泳分析,证实了所获得的抗蛋白抗血清的潜在应用价值。
Preparation of Anti-protein and Anti-mannan Antisera against Fungal Cell Wall by Affinity Chromatography
Iranzo, M., Marcilla, A., Elorza, M. V., Mormeneo, S., and Sentandreu, R. 1994. Preparation of anti-protein and anti-mannan antisera against fungal cell wall by affinity chromatography. Experimental Mycology 18, 159-167. A novel and easy chromatographic method has been developed for the isolation of anti-protein and anti-mannan antisera from a population of polyclonal antibodies obtained against Candida albicans and Yarrowia lipolytica cell wall mannoproteins. The technique is based on the immobilization of mannan (to be used as immunoadsorbent) by Affi-Prep Hz resin after the oxidation of neighboring hydroxyl groups of the polysaccharide with sodium periodate. For Y. lipolytica polyclonal antiserum, a single chromatographic step using the homologous mannan was sufficient to obtain an antiprotein antibody preparation free of antimannan antibodies. For C. albicans, three chromatographic processes using homologous and heterologous mannan were needed to obtain a satisfactory antiprotein antiserum. The potential application of the anti-protein antiserum obtained has been demonstrated by indirect immunofluorescence assays of whole cells and electrophoretic analysis of wall proteins in C. albicans and Saccharomyces cerevisiae .