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Arzneimittelforschung 2001; 51(10): 864-869
DOI: 10.1055/s-0031-1300128
Biotechnology in Drug Research
Editio Cantor Verlag Aulendorf (Germany)

Epitope Specificity of Mouse Immune Response on Short Polypeptides Isolated from Viscum album

Alexander G. Tonevitsky
a   ProBioGen AG, Berlin, Germany
,
Igor Agapov
b   State Scientific Center of Russian Federation “GNllGenetika”, Moscow, Russia
,
Olga Solopova
b   State Scientific Center of Russian Federation “GNllGenetika”, Moscow, Russia
,
Mariya Yurkova
b   State Scientific Center of Russian Federation “GNllGenetika”, Moscow, Russia
c   Institute of Phytochemistry, University of Witten-Herdecke, Witten, Germany
,
Ira Demina
b   State Scientific Center of Russian Federation “GNllGenetika”, Moscow, Russia
,
Karola Pfueller
c   Institute of Phytochemistry, University of Witten-Herdecke, Witten, Germany
,
Uwe Pfueller
c   Institute of Phytochemistry, University of Witten-Herdecke, Witten, Germany
› Author Affiliations
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Publication History

Publication Date:
26 December 2011 (online)

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Summary

A series of mouse monoclonal antibodies to viscotoxins isolated from Viscum album leaves was produced. The interaction of the monoclonal antibodies with different isoforms of viscotoxins was studied by enzymatic immunoassay as well as in the BIAcore (biochemical interaction analysis) system. A highly sensitive test system for the detection of visco-toxin content in biological fluids and commercial extracts from mistletoe, used for pharmacological therapy of malignant tumours, is proposed. Possible anti-gen determinants in viscotoxin structure are proposed. Based on this prediction biotinylated polypeptides corresponding to the most heterogeneous region in the amino acid sequence of viscotoxins were synthesised. The polypeptide APRPTCAK was shown to be the main antigenic epitope for monAbs VH7 and VH10. The test system based on the two monoclonal antibodies VC3 and VH7 has a sensitivity level about 50 ng/ml for viscotoxin A3.

Zusammenfassung

Epitop-Spezifität der Immunantwort der Maus auf kurzkettige Polypeptide isoliert aus Viscum album L.

Es werden monoklonale Antikörper beschrieben, die gegen die Viscotoxine der europäischen Mistel (Viscum album L.) gerichtet sind. Die Interaktion der monoklonalen Antikörper mit den Viscotoxin-Isoformen wurde quantitativ anhand der biomolekularen Interaktionsanalyse (BIA, BIACore) sowie mit Immunoassays (ELISAs) untersucht. Auf dieser Grundlage wird ein empfindliches Testsystem für den Nachweis von Viscotoxinen in biologischen Flüssigkeiten und in kommerziellen Mistelpräparaten, die in der Tumortherapie eingesetzt werden, vorge-schlagen. Eine Vorhersage antigener Determinanten der Viscotoxin-Struktur ermöglichte die Synthese biotinylierter Polypeptide korrespondierend zu den besonders heterogenen Regionen in der Aminosäuresequenz der Viscotoxine. Für die monoklonalen Antikörper VH7 und VH10 ist das Polypeptid APRPTCAK das antigene Hauptepitop. Die Nachweis-grenze eines ELISA-Testsystems mit den monoklonalen Antikörpern VH3 und VH7 beträgt für das Viscotoxin A3 50 ng/ ml.

Key words

Enzyme-linked immunosorbent assay - Monoclonal antibodies - Viscotoxins - Viscum album L.