Quantitation of IgG and IgM Human Anti-Mouse Antibodies (HAMA) Interference in CA 125 Measurements Using Affinity Chromatography
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...
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Published in | Clinical chemistry and laboratory medicine Vol. 36; no. 1; pp. 23 - 28 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
Berlin
Walter de Gruyter
01.01.1998
New York, NY |
Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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Bibliography: | cclm.1998.005.pdf ark:/67375/QT4-G2L8GLG8-F ArticleID:cclm.36.1.23 istex:8F56C57918862D4A2525F48A4D0EB3A978125A66 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 1434-6621 1437-4331 |
DOI: | 10.1515/CCLM.1998.005 |