作者: J. Samuel , G.D. MacLean , A.A. Noujaim , M.R. Suresh , B.M. Longenecker
DOI:
关键词: Antigen 、 H antigen 、 Affinity chromatography 、 MNS antigen system 、 Biochemistry 、 Thomsen-Friedenreich Antigen 、 Immunoassay 、 Radioimmunoassay 、 Chemistry 、 Antibody
摘要: The Thomsen-Friedenreich (TF) antigen is a precursor structure of MN blood group antigens and also expressed by about 90% human carcinomas. immunodominant TF [β-galactosyl(1–3)-α- N -acetylglactosamine] present in cryptic form normal RBC revealed neuraminidase treatment. A murine monoclonal antibody (Mab 49H.8) developed against treated was reactive variety tumors. We have characterized the tumor associated detected this from transitional bladder carcinoma cell line (647V), colon adenocarcinoma (LS174T), pleural effusion fluid breast patient (PE 89). heterologous sandwich radio-immunoassay for using Mab 49H.8 as catcher 125I-peanut agglutinin probe. Detergent extracts 647V LS174T cells, media conditioned culturing these PE 89 were shown to contain assay. specificity capture assay demonstrated its selective inhibition nitrophenyl-β-d-galactoside, phenyl-β-d-galactoside, hapten. Preliminary studies on serum samples showed that 53.7% contained an concentration greater than 200 units/ml whereas 90.9% below units/ml. These shed cells both vitro vivo . sensitive treatment with alkali (0.1 m NaOH 5 h at 37°C) periodate (10 mm sodium 1 room temperature), resistant acidic pH (50 acetate buffer, 4.5, 37°C), could be extracted perchloric acid (0.6 4°C). high molecular weight glycoprotein ( M r >1,000,000) gel filtration chromatography. density estimated 1.35 g/ml cesium chloride gradient centrifugation. isolated combination affinity chromatography overall purification 61,432-fold final recovery 53.2%. Immunoprecipitation detergent extract surface glycoconjugates labeled 3H it approximate 1,000,000. suggest secreted into body fluids mucin-like glycoprotein.