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Monoclonal antibodies against different epitopes of peptide hormones

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Jurzak,  Mirek
Emeritusgroup Physical Chemistry, Max Planck Institute of Biophysics, Max Planck Society;

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Boer,  Rainer
Emeritusgroup Physical Chemistry, Max Planck Institute of Biophysics, Max Planck Society;

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Fritzsch,  Günter
Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society;

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Kojro,  Elzbieta
Emeritusgroup Physical Chemistry, Max Planck Institute of Biophysics, Max Planck Society;

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Fahrenholz,  Falk
Emeritusgroup Physical Chemistry, Max Planck Institute of Biophysics, Max Planck Society;

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Citation

Jurzak, M., Boer, R., Fritzsch, G., Kojro, E., & Fahrenholz, F. (1990). Monoclonal antibodies against different epitopes of peptide hormones. European Journal of Biochemistry, 190(1), 45-52. doi:10.1111/j.1432-1033.1990.tb15543.x.


Cite as: http://hdl.handle.net/21.11116/0000-0007-A242-7
Abstract
In order to produce monoclonal antibodies directed against different epitopes of the neurohypophyseal hormone vasopressin, the hormone was coupled to carrier proteins via photoreactive groups at different positions in the vasopressin sequence: [2‐(4‐azidophenylalanine), 8‐arginine]vasopressin (peptide P1, photoreactive group at position 2) and desamino‐[8‐N6‐(4‐azidophenylamidino)lysine]vasopressin (peptide P2, photoreactive group at position 8) were conjugated to thyroglobulin by flash photolysis. Monoclonal antibodies against these conjugates bound {[3H]8‐arginine}vasopressin with dissociation constants ranging over 40–400 nM. Epitope analysis by means of competitive ELISA showed that the monoclonal antibody obtained with peptide P1 as hapten was directed against the C‐terminal acyclic tripeptide when its conformation was stabilized by interaction with the disulphide‐linked cyclic hexapeptide. In contrast, the epitope analysis of three monoclonal anti‐(peptide P2) antibodies demonstrated that they recognized antigenic determinants in the cyclic hexapeptide ring, mainly the hydrophobic surface formed by Tyr2 and Phe3. Our results suggest that monoclonal antibodies against different epitopes in small peptide hormones can be generated selectively by using photoreactive peptides in such a way that different antigenic sites are exposed in the hapten‐carrier conjugate.