Abstract:
BACKGROUND:Although a variety of animals have been used to produce polyclonal antibodies against antigens, the production of antigen-specific monoclonal antibodies from animals remains challenging. RESULTS:We propose a simple and rapid strategy to produce monoclonal antibodies from a variety of animals. By staining lymph node cells with an antibody against immunoglobulin and a fluorescent dye specific for the endoplasmic reticulum, plasma/plasmablast cells were identified without using a series of antibodies against lineage markers. By using a fluorescently labeled antigen as a tag for a complementary cell surface immunoglobulin, antigen-specific plasma/plasmablast cells were sorted from the rest of the cell population by fluorescence-activated cell sorting. Amplification of cognate pairs of immunoglobulin heavy and light chain genes followed by DNA transfection into 293FT cells resulted in the highly efficient production of antigen-specific monoclonal antibodies from a variety of immunized animals. CONCLUSIONS:Our technology eliminates the need for both cell propagation and screening processes, offering a significant advantage over hybridoma and display strategies.
journal_name
BMC Bioljournal_title
BMC biologyauthors
Kurosawa N,Yoshioka M,Fujimoto R,Yamagishi F,Isobe Mdoi
10.1186/1741-7007-10-80subject
Has Abstractpub_date
2012-09-28 00:00:00pages
80issn
1741-7007pii
1741-7007-10-80journal_volume
10pub_type
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