Cancer Chemotherapy and Pharmacology

, Volume 27, Issue 2, pp 115–120 | Cite as

A highly sensitive enzyme-linked immunosorbent assay for etoposide using β-d-galactosidase as a label

  • Tetsuya Saita
  • Kunio Fujiwara
  • Tsunehiro Kitagawa
  • Masato Mori
  • Katsumi Takata
Original Articles Etoposide, Galactosidase, ELISA

Summary

A highly sensitive enzyme-linked immunosorbent assay (ELISA) for etoposide (EP) was developed, which is capable of accurately measuring as little as 40 pg EP/ml. Anti-EP sera were obtained by immunizing rabbits with EP conjugated with mercaptosuccinyl bovine serum albumin (MS.BSA) usingN-[β-(4-diazophenyl)ethyl]maleimide (DPEM) as a heterobifuntional coupling agent. An enzyme marker was similarly prepared by coupling EP with β-d-galactosidase (β-Gal; EC 3.2.1.23) via DPEM. This ELISA was specific for EP and showed a very slight cross-reactivity with its major metabolite,cis-hydroxy acid of EP (0.91%), but none with 4′-demethylepipodophyllotoxin and drugs commonly used with EP in combination chemotherapy for cancer treatment. The values for EP concentration detected by this assay were comparable with those detected by the highperformance liquid chromatography (HPLC) method. However, the ELISA was about 1,250 times more sensitive in detecting EP at lower concentrations. Using this assay, drug levels were easily determined in the blood and urine of rats for 7 h after i.v. administration of EP at a single dose of 3 mg/kg. Due to its sensitivity and specificity for EP, the ELISA should prove to be a valuable new tool for use in clinical pharmacological studies.

Keywords

Ethyl Albumin Liquid Chromatography Bovine Serum Albumin Serum Albumin 

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Tetsuya Saita
    • 1
  • Kunio Fujiwara
    • 2
  • Tsunehiro Kitagawa
    • 2
  • Masato Mori
    • 1
  • Katsumi Takata
    • 1
  1. 1.Saga Medical SchoolFaculty of Hospital PharmacySagaJapan
  2. 2.Pharmaceutical SciencesNagasaki UniversityNagasakiJapan

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