Breast cyst fluid protein assay

Claim 2

Gross-cystic disease fluid protein-15 (GDCFP-15) essentially free of other components found in human breast gross cystic disease fluid and being further characterized as follows: (a) a glycoprotein having a calculated monomer size of about 15,000 daltons as determined by sodium dodecyl sulfate acrylamide gel analysis; and (b) immunologically not identical to any components of plasma as determined by Ouchterlony analysis; and (c) immunological cross identity with a component of human milk and human saliva; and (d) cleavage with cyanogen bromide provides two peptides one of which is blocked, and one of the peptides has a molecular weight of about 12,500 daltons as determined by sodium dodecyl sulfate acrylamide gel analysis and having the following partial sequence of amino acids: ##STR2##

Claim 3

GCDFP-15 according to claim 1 which is labelled with a unique and detectable label.

Claim 4

The compound of claim 2 wherein said unique and detectable label is a radioactive isotope.

Claim 5

The compound of claim 3 which is 125 I-GCDFP-15.

Claim 6

An antibody specific to the glycoprotein GCDFP-15 according to claim 1.

Claim 7

A process for isolation of Gross cystic disease fluid protein 15 (GDCFP-15) from human breast gross cystic disease fluid which process comprises the following steps in combination: (a) Column chromatograph ultracentrifuged gross cystic disease fluid on Sephadex G-200 to yield peaks of approximately 14,000 and 70,000 molecular size; (b) Hydroxylapatite chromatograph the 70,000 molecular size peak of Step A using sequential buffer applications of 0.01 M, pH 4.8; 0.15 M, pH 4.8; and 0.15 M, pH 8.6, NaH 2 PO 4 buffer; (c) Ion exchange chromatograph the 0.15 M, pH 8.6 eluate from Step B on DEAE-Agarose, pre-equilibrated with 0.05 M NaH 2 PO 4 , pH 4.8 buffer, using sequential buffer applications of 0.05 M, 0.075 M and 0.5 M NaH 2 PO 4 , pH 4.8 buffer; and (d) Recycle the 0.5 M NaH 2 PO 4 , pH 4.8 eluate from Step C through the ion exchange procedure of Step C.

Claim 8

A process for isolation of Gross cystic disease fluid protein 15 (GDCFP-15) from human breast gross cystic disease fluid which process comprises the following steps in combination: (a) Column chromatograph ultracentrifuged gross cystic disease fluid on Sephadex G-200 to yield peaks of approximately 14,000 and 70,000 molecular size; (b) Hydroxylapatite chromatograph the 70,000 molecular size peak of Step A using sequential buffer applications of 0.01 M, pH 4.8; 0.15 M, pH 4.8; and 0.15 M, pH 8.6, NaH 2 PO 4 buffer; (c) Ion exchange chromatograph the 0.15 M, pH 8.6 eluate from Step B on DEAE-Agarose, pre-equilibrated with 0.05 M NaH 2 PO 4 , pH 4.8 buffer, using sequential buffer applications of 0.05 M, 0.075 M and 0.5 M NaH 2 PO 4 , pH 4.8 buffer; and (d) Recycle the 0.5 M NaH 2 PO 4 , pH 4.8 eluate from Step C through the ion exchange procedure of Step C.