Herstellung von katalytischen monoklonalen Antikörpern

Generation of catalytic monoclonal antibodies

by Farrokh Djalali Bazzaz
Doctoral thesis
Date of Examination:2000-10-31
Date of issue:2001-01-15
Advisor:Prof. Dr. Johann Hinrich Peters
Referee:Prof. Dr. Ulrich Grossbach
Referee:Prof. Dr. Friedrich-Wilhelm Schürmann
Persistent Address: http://dx.doi.org/10.53846/goediss-675

 

 

Files in this item

Name:diss.pdf
Size:848.Kb
Format:PDF
Description:Dissertation

The following license files are associated with this item:

Abstract

English

Utilizing the similar binding characteristics of substrates to enzymes and antigens to antibodies, respectively, antibodies with catalytic activities have been created since 1986. We have succeeded in developing the first monoclonal antibody that catalyzes a cyclic reaction to form vitamin E in a stereoselective manner. So far, vitamin E can chemically be synthesized in a mixture of eight stereoisomers which have only a limited applicability for human use. Haptens and substrates have been synthesized with predefined entantiomeric key structures representing the chroman ring system of vitamin E. Mice strains with a higher property of forming catalytic antibodies were immunized with KLH conjugated hapten. The monoclonal antibodies developed were prescreened for affinity and thereafter for their catalytic activity. For this purpose a novel cat-ELISA was designed by indirectly immobilizing the antibody to the solid phase. This enabled us to recover substrate as well as product in a soluble form and to further chemically characterize them. The cat-reaction was run in pure methanol which may be the first example of a non-aqueous antibody reaction. The cat-mAb Doro-1 catalyses the syntheses of alpha-tocopherol with 5% product and an ee-value up to 42% measured by TLC and HPLC, (respectively).Moreover we show that a specific monoclonal antibody against vitamin E (mAb Ing-1, an IgG2a, with kappa light chain and a dissociation constant of KD= 6.72 +/- 2.10 x 10-9 M) have been generated and used for low level detection of vitamin E.
Keywords: catalytic antibody; cat-ELISA; Vitamin E; Hapten
 
Statistik