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Production d'anticorps monoclonaux dirigés contre des peptides de 3, 4 et 6 acides aminés identifiés dans le sérum humain : essai de mise au point d'un dosage immunologique du peptide HWESAS

Abstract : Human plasma contains low molecwar weight growth factors potentiating certain in vitro biological effects of insulin-like growth factors. Recently three peptides were identified with HWESAS, HWE and WGHE as respective structures. Although it is difficult to product monoclonal antibodies directed against peptides characterized by a structure consisting of an amino acid number below 8, this study describes the production of such antibodies for peptides having 6, 4, or 3 residus. The mice immunizations were performed with homologous synthetic peptides coupled to a maleimide activated keyhole limpet hemocyanin and the hybridomas were prepared by standard techniques. By using an inhibition test, the specificity of monoclonal antibodies was deterrninated : they cross-reacted with original peptides as well as derivated peptides only when C terminus was not modified. Concerning the affinity, measured by non competitive enzyme immunoassay, the values range from 0,89 to 3,89 108 M-1. In addition, an ELISA was developped with antibodies directed against HWESAS to measure the serie concentration of this peptide in humans. The levels of peptide observed in healthy subjects were about 2µg/ml, they were dramatically decreased in chronic renal failure but norrnalized after a successful renal transplantation. These findings suggest that the kidney may play an important role in the production and/or metabolism of this hexapeptide. Such monoclonal antibodies will be useful tools for elucidation of low molecular weight growth factor production leading to a better understanding of their physiological and pathological significances.
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http://docnum.univ-lorraine.fr/prive/SCD_T_1999_0331_VIN_CAPIAUMONT.pdf
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https://hal.univ-lorraine.fr/tel-01747101
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Submitted on : Thursday, March 29, 2018 - 10:51:02 AM
Last modification on : Thursday, January 14, 2021 - 11:35:17 AM

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Josette Vin-Capiaumont. Production d'anticorps monoclonaux dirigés contre des peptides de 3, 4 et 6 acides aminés identifiés dans le sérum humain : essai de mise au point d'un dosage immunologique du peptide HWESAS. Médecine humaine et pathologie. Université Henri Poincaré - Nancy 1, 1999. Français. ⟨NNT : 1999NAN10331⟩. ⟨tel-01747101⟩

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