Original Paper

An Enzyme-Linked Immunosorbent Assay for the Measurement of Circulating Antibodies to Polymorphic Epithelial Mucin (MUC1)
Silvia von Mensdorff-Pouilly^a, Maia M. Gourevitch^a, Peter Kenemans^a, Albert A. Verstraeten^a, Gerard J. van Kamp^b, Astrid Kok^b, Kees van Uffelen^b, Frank G.M. Snijdewint^a, Marinus A. Paul^d, Sybren Meijer^c, Joseph Hilgers^a

Departments of
^a Obstetrics and Gynaecology
^b Clinical Chemistry
^c Surgery, Academic Hospital Vrije Universiteit, Amsterdam, and
^d Department of Surgery, Zuiderziekenhuis, Rotterdam, The Netherlands

Address of Corresponding Author

Tumor Biol 1998;19:186-195 (DOI: 10.1159/000030006)

  Key Words

• ELISA
• MUC1
• PEM
• Autoantibodies
• CA 15-3
• Breast cancer

  Abstract

Introduction: About one-third of breast and ovarian carcinoma patients have circulating antibodies reactive with polymorphic epithelial mucin (MUC1), either free or bound to immune complexes. While the presence of these immune complexes has prognostic significance in breast cancer patients, the significance of free MUC1 antibodies is less clear. The objective of this study was to develop a reliable assay for the accurate determination of circulating free antibodies to MUC1. Material and Methods: We developed an enzyme-linked immunosorbent assay (ELISA) (PEM.CIg) employing a 60 mer peptide (a triple tandem repeat sequence of the MUC1 peptide core) conjugated to bovine serum albumin and peroxidase-labeled antihuman immunoglobulin G or M antibodies. The assay was standardized and its analytical performance evaluated. A total of 492 serum samples were obtained from 40 healthy men, from 201 healthy women (including 55 women without a history of pregnancy and 45 pregnant women), and (before primary treatment) from 62 benign breast tumor patients and 190 breast cancer patients. MUC1 serum levels were determined with commercial CA 15-3 tests. Results: Circulating antibodies to MUC1 are present both in healthy subjects and in breast cancer patients. The within- and between-assay coefficients of variation were, respectively, 2 and 12% for the IgG determinations and 1.2 and 3% for the IgM determinations. Correlation coefficients for serially diluted serum samples ranged from 0.9998 to 0.9920 for IgG and from 0.9996 to 0.9818 for IgM determinations. The reactivity of serum samples was partially blocked by the addition of various MUC1 peptides and by MUC1 mucin. The inhibiting effect of modified 60 mer peptides suggests the presence of antibodies directed to more than one epitope. Conclusions: The PEM. CIg assay is a reliable ELISA for measuring free MUC1 antibodies in serum. We are in the process of relating the results obtained in the breast cancer group to disease outcome to evaluate its prognostic significance. In addition, the assay may become a useful tool for vaccine therapy monitoring.

  Author Contacts

Silvia von Mensdorff-Pouilly, MD, Department of Obstetrics and
Gynecology, Academic Hospital Vrije Universiteit, De Boelelaan 1117
NL-1081 HV Amsterdam (The Netherlands)
Tel. +3120 4444829, Fax +3120 4444818,
E-Mail s.vonmensdorff@azvu.nl

  Article Information

Received: Received: August 15, 1997
Accepted after revision: October 16, 1997
Number of Print Pages : 10
Number of Figures : 4, Number of Tables : 1, Number of References : 40