Rapid production of antigen-specific monoclonal antibodies from a variety of animals

Autor: Masaharu Isobe, Rika Fujimoto, Fuminori Yamagishi, Megumi Yoshioka, Nobuyuki Kurosawa
Rok vydání: 2012
Předmět:
Physiology
ER-tracker
Cell Separation
Plant Science
Endoplasmic Reticulum
Epitopes
Antibody Specificity
Structural Biology
Antigen specific
Cell separation
Insulin
rat
lcsh:QH301-705.5
Phylogeny
Immunoassay
medicine.diagnostic_test
biology
Agricultural and Biological Sciences(all)
Methodology Article
Antibodies
Monoclonal
Flow Cytometry
MAGrahd
single cell
antigen-specific monoclonal antibody
Antibody
General Agricultural and Biological Sciences
Biotechnology
medicine.drug_class
Plasma Cells
FACS
rabbit
Monoclonal antibody
General Biochemistry
Genetics and Molecular Biology
Flow cytometry
TS-jPCR
Antigen
ERIAA
medicine
Animals
Humans
Cell Lineage
human
Ecology
Evolution
Behavior and Systematics
Fluorescent Dyes
Biochemistry
Genetics and Molecular Biology(all)
Cell Biology
Virology
Epitope mapping
lcsh:Biology (General)
Polyclonal antibodies
Immunoglobulin G
Immunology
biology.protein
Biomarkers
Epitope Mapping
guinea pig
Developmental Biology
Zdroj: BMC Biology
BMC Biology, Vol 10, Iss 1, p 80 (2012)
ISSN: 1741-7007
DOI: 10.1186/1741-7007-10-80
Popis: 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.
Databáze: OpenAIRE
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