Premium
Microimmunofluorescence Using Terasaki Plates and Direct Plate Freezing Method—Rapid and Reliable Screening System of Hybridomas
Author(s) -
Nakagawa Shoko,
Tsuji Yoshiyuki,
Nishiura Haruhiko,
Isojima Shinzo
Publication year - 1986
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1986.tb03045.x
Subject(s) - biology , glutaraldehyde , monoclonal antibody , antibody , microbiology and biotechnology , staining , chromatography , immunology , chemistry , genetics
Microimmunofluorescence using Terasaki plates and a direct plate freezing method were combined for effective screening of hybridoma supernatants. The microplates, in which the fused cells (myeloma and spleen cells) were cultured and hybridoma colonies were growing, were frozen after harvest of supernatants and saved at −80 C for several weeks without affecting antibody production ability of hybridomas. Microimmunofluorescence was performed in Terasaki plates on which target cells were attached by poly‐l‐lysine and glutaraldehyde or by short time culture of the cells in Terasaki plates. The direct plate freezing method prevented initial hybridoma cells from changes or disappearance of antibody productions during screening of hybridoma supernatants; the microimmunofluorescence staining method permits fast and detailed estimation of specificity of antibodies of hybridomas by saving time and minimal consumption of supernatant for checking. The combination of these two methods is a powerful tool for obtaining desired monoclonal antibodies.