Generating monoclonal antibodies to neuronal antigens.
Methods for generating monoclonal antibodies directed to the functional sites of neuronal antigens are reviewed. These methods include optimal antigen preparation and presentation as well as selective targeting and manipulation of the antigenic response. We describe our use of the immunosuppressant drug, cyclophosphamide, to produce a selective immune response to rare, poorly immunogenic, or actively suppressed antigens. These techniques allow us to generate antibodies to the functional sites of neuronal antigens, such as cell surface molecules. Such antibodies are directed to complex carbohydrates, proteins, protein complexes and glycolipids that form the active site of neuronal antigens. We can use these antibodies in the molecular dissection of functional active sites that are inaccessible to genetic manipulation. These techniques favor the generation of antibodies that can be used to understand and manipulate neuronal cellular activity.
Department of Neurobiology, Duke University Medical Center, Durham, NC 27710, USA. riggottm@ohsu.edu
M J
MJ
Riggott
W D
WD
Matthew
D000818:Animals; D000911:Antibodies, Monoclonal / Q000187:drug effects; D000941:Antigens / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects*; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism; D003520:Cyclophosphamide / Q000494:pharmacology*; D007150:Immunohistochemistry / Q000379:methods*; D051381:Rats
D000911:Antibodies, Monoclonal; D000941:Antigens; D003520:Cyclophosphamide