H. Vollmers, E. Woźniak, Ewa Stepien-Bötsch, U. Zimmermann, H. Müller-hermelink
{"title":"一种快速从大众培养中纯化人IgM单克隆的方法。","authors":"H. Vollmers, E. Woźniak, Ewa Stepien-Bötsch, U. Zimmermann, H. Müller-hermelink","doi":"10.3233/HAB-1996-7105","DOIUrl":null,"url":null,"abstract":"We describe in this paper the rapid and cheap purification of human immunoglobulin M from hybridoma supernatant of mass culture. The method consists of two steps: 1. concentration of supernatant by ultrafiltration and 2. dialysis against distilled water, pH 6.4. To produce the supernatant, the hybridomas are grown in RPMI media with fetal calf serum (10% FCS) in 250 ml flasks under normal tissue culture conditions. More than 14 mg IgM can be nearly selectively purified within 6 h from 5 l of antibody containing hybridoma supernatant. Most of the IgM molecules stay in a penta- and monomeric form and are positive in physiological and immunohistochemical studies.","PeriodicalId":77166,"journal":{"name":"Human antibodies and hybridomas","volume":"7 1 1","pages":"37-41"},"PeriodicalIF":0.0000,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3233/HAB-1996-7105","citationCount":"12","resultStr":"{\"title\":\"A rapid method for purification of monoclonal human IgM from mass culture.\",\"authors\":\"H. Vollmers, E. Woźniak, Ewa Stepien-Bötsch, U. Zimmermann, H. Müller-hermelink\",\"doi\":\"10.3233/HAB-1996-7105\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"We describe in this paper the rapid and cheap purification of human immunoglobulin M from hybridoma supernatant of mass culture. The method consists of two steps: 1. concentration of supernatant by ultrafiltration and 2. dialysis against distilled water, pH 6.4. To produce the supernatant, the hybridomas are grown in RPMI media with fetal calf serum (10% FCS) in 250 ml flasks under normal tissue culture conditions. More than 14 mg IgM can be nearly selectively purified within 6 h from 5 l of antibody containing hybridoma supernatant. Most of the IgM molecules stay in a penta- and monomeric form and are positive in physiological and immunohistochemical studies.\",\"PeriodicalId\":77166,\"journal\":{\"name\":\"Human antibodies and hybridomas\",\"volume\":\"7 1 1\",\"pages\":\"37-41\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1996-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.3233/HAB-1996-7105\",\"citationCount\":\"12\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Human antibodies and hybridomas\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.3233/HAB-1996-7105\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Human antibodies and hybridomas","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.3233/HAB-1996-7105","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
A rapid method for purification of monoclonal human IgM from mass culture.
We describe in this paper the rapid and cheap purification of human immunoglobulin M from hybridoma supernatant of mass culture. The method consists of two steps: 1. concentration of supernatant by ultrafiltration and 2. dialysis against distilled water, pH 6.4. To produce the supernatant, the hybridomas are grown in RPMI media with fetal calf serum (10% FCS) in 250 ml flasks under normal tissue culture conditions. More than 14 mg IgM can be nearly selectively purified within 6 h from 5 l of antibody containing hybridoma supernatant. Most of the IgM molecules stay in a penta- and monomeric form and are positive in physiological and immunohistochemical studies.