Monoclonal antibody M3 used in tissue polypeptide-specific antigen assay for the quantification of tissue polypeptide...

Recently, a new 'specific tissue polypeptide antigen (TPA)' test was introduced and designated tissue polypeptide-specific antigen (TPS); it is based on the monoclonal antibody (MAb) anti-TPS, M3. We have tested the specificity of this antibody by immunocyto- and immunohistochemistry, gel electrophoresis and immunoblotting. MAb M3 bound to intermediate filaments of epithelial cells and revealed a staining pattern identical to cytokeratin (CK) 18-specific MAb (DE-K18) on tissue sections of various human tissues. On immunoblots of proteins extracted from various epithelial cell lines, M3 reacted with a 45-kD protein corresponding to CK18, and on immunoblots of proteins isolated from MCF-7 culture fluid M3 stained three bands, 45, 33 and 29 kD. The same bands were stained with CK18-specific MAb, indicating that they represent CK18 and its degradation products. TPA, used as a tumor marker in clinical diagnoses and follow-up, was shown to be a degradation product of CK 8, 18 and 19. In contrast to TPA, MAb M3 did not stain CK8 and CK19 present on immunoblots.