biostudies-literatureSzafran ATNICHD NIH HHSDiana Helis Henry Medical Research FoundationNIDDK NIH HHSNIEHS NIH HHSNIEHSCenter of Excellence in Environmental HealthBaylor College of MedicineNCI NIH HHSNIH75-84https://www.ebi.ac.uk/biostudies/studies/S-EPMC4891975biostudies-literatureUnknown96Understanding the properties and functions of complex biological systems depends upon knowing the proteins present and the interactions between them. Recent advances in mass spectrometry have given us greater insights into the participating proteomes, however, monoclonal antibodies remain key to understanding the structures, functions, locations and macromolecular interactions of the involved proteins. The traditional single immunogen method to produce monoclonal antibodies using hybridoma technology are time, resource and cost intensive, limiting the number of reagents that are available. Using a high content analysis screening approach, we have developed a method in which a complex mixture of proteins (e.g., subproteome) is used to generate a panel of monoclonal antibodies specific to a Methods (San Diego, Calif.)Use of HCA in subproteome-immunization and screening of hybridoma supernatants to define distinct antibody binding patterns.PMC4891975ES023512-01P30CA125123P30 HD007495T32 HD007495CA125123P30 CA125123U54 HD007495K12 DK083014K12DK0083014HD007495DK56338P30 DK056338P30 ES0235121R01ES023206-01R01 ES023206Nickerson JAMancini MAMancini MGEdwards DPSzafran ATfalseUse of HCA in subproteome-immunization and screening of hybridoma supernatants to define distinct antibody binding patterns.Understanding the properties and functions of complex biological systems depends upon knowing the proteins present and the interactions between them. Recent advances in mass spectrometry have given us greater insights into the participating proteomes, however, monoclonal antibodies remain key to understanding the structures, functions, locations and macromolecular interactions of the involved proteins. The traditional single immunogen method to produce monoclonal antibodies using hybridoma technology are time, resource and cost intensive, limiting the number of reagents that are available. Using a high content analysis screening approach, we have developed a method in which a complex mixture of proteins (e.g., subproteome) is used to generate a panel of monoclonal antibodies specific to a 2016-01-01T00:00:00Z2016 Mar2025-04-19T03:07:41.977Z2019-03-27T02:15:15ZS-EPMC48919752652197610.1016/j.ymeth.2015.10.021