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International Journal of Legal Medicine

, Volume 104, Issue 3, pp 149–152 | Cite as

Production and characterization of a monoclonal antibody to ABH-carrying α2-seminoglycoprotein for ABO grouping of semen by ELISA

  • T. Kishida
  • Y. Tamaki
  • R. Tsuda
  • N. Naharaha
  • Y. Katsumata
  • N. Kimura
Original Articles

Summary

BALB/c mice were immunized with α2-seminoglycoprotein (A2SGP), the lymph node cells were fused with P3U1 myeloma cells and cultured by the conventional technique. Four antibody-producing hybridoma clones were established and antibody-containing ascitic fluid obtained. The antibody was directed to the protein backbone of A2SGP and not to ABH antigenic determinants and did not cross-react with saliva or vaginal secretions. When tested in an indirect ELISA the anti-A2SGP antibody had a titer of 512000. The anti-A2SGP was used in a capture ELISA (or sandwich ELISA) in which wells were coated with this antibody to capture A2SGP in semen, and the captured A2SGP was detected with anti-A, anti-B or anti-H-peroxidase conjugate and peroxidase-labeled second antibody. This ELISA allowed correct ABO grouping even of 1:12800 or higher dilutions of semen. When the ELISA was applied to ABO grouping of seminal fluids mixed with vaginal secretions only the seminal ABH antigens could be detected. The results strongly suggest the potential usefulness of monoclonal anti-A2SGP in the investigation of rape cases.

Key words

Personal identification ABO Semen Monoclonal anti-α2-seminoglycoprotein ELISA 

Zusammenfassung

BALB/c-Mäuse wurden mit α2-Seminoglykoprotein (A2SGP) immunisiert; die Lymphknotenzellen wurden fusioniert mit P3U1 Myelomzellen und mit konventioneller Technik kultiviert. 4 Antikörperproduzierende Hybridoma-Klone wurden etabliert und Antikörper-enthaltende Ascitesflüssigkeit gewonnen. Der Antikörper war gegen das Protein-Rückgrat von A2SGP gerichtet und nicht gegen die ABO-Antigen-Determinanten und er reagierte nicht mit Speichel oder Scheidensekret. Im indirekten ELISA-Test hatte der Anti-A2SGP-Antikörper einen Titer von 512000. Der Anti-A2SGP-Antikörper wurde getestet in einem „Capture-ELISA” (oder „Sandwich-ELISA”), in welchem Vertiefungen mit diesem Antikörper beschichtet wurden, um A2SGP aus Spermaflüssigkeit zu binden und wurde dann mit einem zweiten Anti-A, Anti-B und Anti-H-Antikörper detektiert, welcher mit Peroxydase konjugiert oder markiert war. Dieser ELISA-Test erlaubte eine korrekte ABO-Bestimmung, selbst bei 1:12800 oder höheren Verdünnungen der Spermaflüssigkeit. Bei Gemischen von Spermaflüssigkeit mit Vaginalsekret war der ELISA-Test zum ABO-Nachweis imstande, nur die ABH-Antigene der Spermaflüssigkeit zu detektieren. Die Ergebnisse suggerieren eindrucksvoll die potentiellen Möglichkeiten des monoklonalen Anti-A2SGP-Antikörpers für die Untersuchung von Vergewaltigungsfällen.

Schlüsselwörter

Spermaflüssigkeit Monoklonales Anti-α2-Seminoglykoprotein ABO 

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References

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Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • T. Kishida
    • 1
  • Y. Tamaki
    • 1
  • R. Tsuda
    • 2
  • N. Naharaha
    • 3
  • Y. Katsumata
    • 4
  • N. Kimura
    • 2
  1. 1.Department of Forensic MedicineMedical College of OitaOitaJapan
  2. 2.Department of Legal MedicineKurume University School of MedicineKurumeJapan
  3. 3.Department of Obstetrics and GynecologyMedical College of OitaOitaJapan
  4. 4.Department of Legal MedicineNagoya University School of MedicineNagoyaJapan

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