Alemtuzumab: Validation of a sensitive and simple enzyme-linked immunosorbent assay

Iman Jilani, Michael Keating, Francis J. Giles, Susan O'Brien, Hagop M. Kantarjian, Maher Albitar*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

24 Scopus citations


Alemtuzumab (MabCampath®) is a humanized rat monoclonal antibody that targets the CD52 antigen. It has been approved for the treatment of patients with resistant chronic lymphocytic leukaemia (CLL). Measuring plasma/serum levels of alemtuzumab is important for optimizing the dosing and scheduling of therapy; however, current assays in serum or plasma, based on the capture of alemtuzumab using CD52, are complicated and difficult to adapt for high throughput testing. We developed a simple sandwich enzyme-linked immunosorbent assay (ELISA) to measure alemtuzumab that takes advantage of the remaining rat sequence in alemtuzumab. Using specific anti-rat immunoglobulin (Ig) antibodies (absorbed against human Ig), alemtuzumab levels were measured in the serum and plasma of patients treated with alemtuzumab. Levels were similar between plasma and serum samples, in fresh samples and samples stored at 4 °C for 24 h, but were significantly lower in samples stored at room temperature for 24 h. The assay was successfully used to determine serum alemtuzumab pre- and post-treatment. This assay is simple and adaptable for high throughput testing, with a limit of detection of 0.05 μg/ml and a coefficient of variation of ±12.5%. No false positivity was observed in >200 samples tested. This validated assay should help optimize the dosing and scheduling of alemtuzumab therapy. The underlying principles are also applicable to the measurement of other humanized antibodies using an appropriate anti-Ig.

Original languageEnglish (US)
Pages (from-to)1255-1262
Number of pages8
JournalLeukemia Research
Issue number12
StatePublished - Dec 2004


  • Alemtuzumab
  • CLL
  • Humanized
  • MabCampath

ASJC Scopus subject areas

  • Hematology
  • Oncology
  • Cancer Research


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