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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Schlattmann, Peter / Tate, Jillian R. / Tsongalis, Gregory J.

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Quantitation of IgG and IgM Human Anti-Mouse Antibodies (HAMA) Interference in CA 125 Measurements Using Affinity Chromatography

Norbert P. Koper / Chris M. G. Thomas / Leon F. A. G. Massuger / Martin F. G. Segers / André J. Olthaar / André L. M. Verbeek

Citation Information: Clinical Chemistry and Laboratory Medicine. Volume 36, Issue 1, Pages 23–28, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.1998.005, June 2005

Publication History

Published Online:
2005-06-01

Abstract

Currently no available immunoassay system offers complete protection against spuriously elevated or lowered results due to interference by Human Anti-Mouse Antibodies (HAMA). Although routine use of chromatography procedures is not an acceptable option because of the extra cost and workload involved, such a procedure would be highly desirable to ensure accurate immunoassay results. The present report describes a relatively simple affinity chromatography procedure using a HiTrap Protein G column to isolate immunoglobulin G (IgG) HAMA, followed by a HiTrap N-hydroxy-succinimide(NHS)-activated column coupled to goat-anti human immunoglobulin M (IgM) to bind IgM HAMA. To examine the usefulness of this purification procedure we determined CA 125 in forty serum samples prior to and following chromatography. Pre- and post-injection samples were obtained from 20 patients injected with 1 mg of 111In-Iabelled murine OC 125 F(ab′)2 fragments in an immunoscintigraphy study.

It is shown that this analytical procedure provides a technique to determine the extent and the nature of the existing HAMA interference in samples of patients after in vivo use of monoclonal antibodies for diagnostic or therapeutic purposes. The procedure can also contribute to the clarification of clinically discordant CA 125 results. Finally, the availability of such a procedure in the clinical laboratory provides an opportunity to test the robustness of newly developed immunoassay systems towards HAMA interference.

Citing Articles

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[1]
Norbert P. Koper, Leon F.A.G. Massuger, Chris M.G. Thomas, Cornelis Beyer, and Marinus J. Crooy
European Journal of Obstetrics & Gynecology and Reproductive Biology, 1999, Volume 86, Number 2, Page 203
[2]
Angèle L.M. Oei, Fred C.G.J. Sweep, Leon F.A.G. Massuger, André J. Olthaar, and Chris M.G. Thomas
Gynecologic Oncology, 2008, Volume 109, Number 2, Page 199
[3]
Chris MG Thomas, Leon FAG Massuger, and Hans MWM Merkus
The Lancet, 2000, Volume 355, Number 9216, Page 1725
[4]
A. K. Knight, T. Bingemann, L. Cole, and C. Cunningham-Rundles
Clinical and Experimental Immunology, 2005, Volume 141, Number 2, Page 333

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