Use of enantioselective antibodies in analyt. techniques

Subject of this project is the production of enantioselective antibodies and their application for chiral recognition in immunoassay techniques and separation methods. Goal of this work is the development of highly sensitive and selective methods for enantiomer purity checks. It is well established that the pharmacological activities of drugs are restricted to one of the enantiomers, while the other one may exhibit unwanted or even toxic side effects. Albumine conjugates of the hapten enantiomers (ß-blockers) will be prepared for antibody production in rabbits. After purification, these antibodies will be first checked for their chiral recognition ability by ELISA. The antibodies should respond only to the enantiomer against which they were raised. Based on these results, enantioselective flow-injection and sequential-injection immunoassays with chemiluminescence detection will be developed which should enable the detection of impurities of the inactive enantiomer in the femtomole range. The main target of this project will be the development of immuno affinity phases containing immobilized antibodies for chiral separation by nano-HPLC. Besides packed columns on silica gel basis, also monolithic columns will be prepared using an in-situ copolymerization process of the antibodies with methacrylamide monomers. Phases on this basis will be applied also in capillaries for capillary electrochromatography and enantioselective extraction approaches. In addition, the stereoselective interactions will be also investigated using capillary zone electrophoresis.

Subject of this project is the production of enantioselective antibodies and their application for chiral recognition in immunoassay techniques and separation methods. Goal of this work is the development of highly sensitive and selective methods for enantiomer purity checks. It is well established that the pharmacological activities of drugs are restricted to one of the enantiomers, while the other one may exhibit unwanted or even toxic side effects. Albumine conjugates of the hapten enantiomers (ß-blockers) will be prepared for antibody production in rabbits. After purification, these antibodies will be first checked for their chiral recognition ability by ELISA. The antibodies should respond only to the enantiomer against which they were raised. Based on these results, enantioselective flow-injection and sequential-injection immunoassays with chemiluminescence detection will be developed which should enable the detection of impurities of the inactive enantiomer in the femtomole range. The main target of this project will be the development of immuno affinity phases containing immobilized antibodies for chiral separation by nano-HPLC. Besides packed columns on silica gel basis, also monolithic columns will be prepared using an in-situ copolymerization process of the antibodies with methacrylamide monomers. Phases on this basis will be applied also in capillaries for capillary electrochromatography and enantioselective extraction approaches. In addition, the stereoselective interactions will be also investigated using capillary zone electrophoresis.