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A M_r 43,000 Epidermal Growth Factor-related Protein Purified from the Urine of Breast Cancer Patients

Central Institute of Molecular Biology [K. E., R. G.], Central Institute of Cancer Research [A. G., J. F.], Academy of Sciences of the German Democratic Republic, 1115 Berlin-Buch, German Democratic Republic; and Centralsygehuset Hillerød, Department of Clinical Chemistry, 3400 Hillerød, Denmark [E. N.]

A tumor-associated epidermal growth factor (EGF)-like activity was detected in the urine of breast cancer patients by means of an EGF radioreceptor assay and an anchorage-independent growth assay. The clonogenic growth factor activity of pooled void volume eluate fractions from a Bio-Gel P-30 column was completely neutralized by an anti-human epidermal growth factor antiserum but not by an anti-transforming growth factor antiserum. This activity was determined in the urine of 71 breast cancer patients. A statistically significant correlation was found between EGF-like clonogenic activity and axillary lymph node status, tumor size, stage of disease, and grade of differentiation of the primary tumor. The Bio-Gel P-30 void volume fraction was used to purify the EGF-related polypeptide to apparent homogeneity by subsequent binding to and elution from A431 cells followed by isoelectric focusing. A polypeptide of a pI of approximately 3.4 was identified to be related to EGF by neutralization and immunoprecipitation experiments with anti-human epidermal growth factor antisera. This polypeptide migrated as a single band of M_r 43,000 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

¹ Supported by a grant from the Danish Cancer Society (89-060).

² To whom requests for reprints should be addressed, at Institute of Molecular Biology, Academy of Sciences, R.-Roessle-Strasse 10, Berlin-Buch 1115, German Democratic Republic.

Received 2/14/89. Revised 8/ 7/89. Revised 10/ 2/89. Accepted 10/13/89.