Isolation and partial characterization of a soluble oncofetal antigen from murine and human amniotic fluids.
A soluble (cell-free) oncofetal antigen (OFA) was detected in murine and human amniotic fluids by immunostaining with the murine monoclonal antibody (MAb 115) produced by syngeneic immunization with mid-gestational mouse fetal cells. OFA was purified from the amniotic fluids by ammonium sulfate precipitation at 30-70% saturation, followed by successive gel chromatography of the OFA-containing fraction on Sephacryl-S300 HR, Q- and S-Sepharoses and lentil lectin agarose. The fraction eluted from the lentil lectin column gave a single band on SDS-PAGE of the same molecular weight as the membrane-bound OFA found on both fetal and tumor tissues of humans and several rodents. Both soluble and membrane-bound OFAs share several chemical characteristics, including binding to lentil lectin and wheat-germ agglutinin, molecular weight (44 kDa) and pI (6.8). Mild periodate oxidation of OFA did not affect its binding to MAb 115 in an enzyme-linked immunosorbent assay, indicating that the reactive epitope is a peptide.
Barsoum
A L
AL
Department of Microbiology and Immunology, College of Medicine, University of South Alabama, Mobile 36688.
Coggin
J H
JH
D000653:Amniotic Fluid / Q000276:immunology*; D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D002853:Chromatography, Liquid / Q000379:methods; D004586:Electrophoresis / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005333:Fetus / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D010504:Periodic Acid
D000951:Antigens, Neoplasm; C031192:oncofetal antigens; D010504:Periodic Acid; C009288:metaperiodate