Core D: Monoclonal Core: (Diamond Director) This Program Project addresses the impact of antibodies to the NMDAR on adult and fetal brain. The unifying hypothesis is that antibody specificity determines the effect of the antibody on brain tissue. Project 1, 2 and 4 all utilize a panel of monoclonal anti-NMDAR antibodies. These will need to be produced in large quantity. The Monoclonal Antibody Core will produce both human and murine antibodies and will test these for preservation of antigen-binding. The Core will produce antibody at high concentration as ascites or in miniPERM apparati. The IgG will be purified on Protein G columns. The Core will generate human IgG from the immunoglobulin variable regions of selected phage or peripheral blood B cells. The Core will generate NR2A and NR2B fragments containing the extracellular amino acids for use in ELISAs. This facility will be cost-effective and will maintain quality control for 3 projects in the Program. This represents an efficient approach to the generation of reagents to be used by multiple investigators and one that maximizes the production of high quality reagents.
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