Modification of Antibody Affinity by Directed Mutagenesis
{'Code': '08060200', 'Directorate': {'LongName': 'Direct For Biological Sciences'}, 'Division': {'LongName': 'Division of Cellular Biosciences'}}
{'SignBlockName': 'Maryanna P. Henkart'}
This project will utilize a T7 expression vector for the                        production of anti-dansyl VH protein in E. coli.  The VH proteins               associated with two anti-dansyl hybridomas have been purified                   from bacterial extracts and recombined with homologous light                    chains to yield fully active VH-L products.  These VH genes will                be subjected to site-directed mutagenesis with single or paired                 primers to provide mutated VH proteins with replacements in the                 hypervariable regions.  The mutant VH proteins will be recombined               with L chains and the purified products characterized with                      respect to affinity by resonance energy transfer, a method                      established in this laboratory.  In addition, the CDR3 segments                 of high affinity and low affinity VDJ genes will be interchanged                to evaluate the role of CDR3 in specific reactivity, particularly               with germ-line (IgM) antibodies.                                                                                                                                The goal of this research is development of a general method to                 manipulate the affinity of monoclonal antibodies for their                      ligands using techniques of genetic engineering.  If this project               succeeds the method will have many practical as well as basic,                  scientific applications.
{'FirstName': 'Fred', 'LastName': 'Karush', 'EmailAddress': None, 'StartDate': '08/15/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'University of Pennsylvania', 'CityName': 'Philadelphia', 'ZipCode': '191046205', 'PhoneNumber': '2158987293', 'StreetAddress': 'Research Services', 'CountryName': 'United States', 'StateName': 'Pennsylvania', 'StateCode': 'PA'}
{'Code': '1117', 'Text': 'CELLULAR PHYSIOLOGY'}
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1988&All=true', 'xml': '8801887.xml'}