Monoclonal Antibodies Against Afp: Application in Immunoassay and Affinity Chromatography

Abstract

Twenty-two hybridoma clones which produced monoclonal antibodies to AFP were established by the fusion of myeloma cells and spleen cells of mice immunized with AFP. Antibodies secreted in the ascites of mice in which the tumor was transplanted intraperitoneally were used in the radioimmunoassay of AFP by the sandwich method; these antibodies were also used in the immunoaffinity purification of AFP. The results indicated that some antibodies had several advantages over the conventional polyclonal antibodies from antiserum in these procedures.

Establishment of a monoclonal antibody recognizing an antigenic site common to Clostridium botulinum type B, C1, D, and E toxins and tetanus toxin

“…Purification of monoclonal antibodies. Immunoglobulin was purified from ascites fluids as described previously (7). Ascites fluids (4 ml) were dialyzed against 0.1 M Tris hydrochloride (pH 7.4) and subjected to column chromatography on DEAE-cellulose (1.6 by 10 cm; DE52; Whatman, Inc., Maidstone, Kent, England) equilibrated with the buffer described above.…”

Modulation of T Cell Function by Alpha-Fetoprotein: An in vivo Study on Porcine Thyroid Peroxidase-Induced Experimental Autoimmune Thyroiditis in Transgenic Mice Producing Human Alpha-Fetoprotein

“…Human AFP was purified from ascites fluid of patients with hepatocellular carcinoma by 70% saturated ammonium sulfate precipitation and followed by antibody affinity column chromatography using mouse monoclonal antibody (MAb) against human AFP (Y-2), as described elsewhere [17].…”

Analysis of Epitopes of Mouse Monoclonal Antibodies against Human Alpha-Fetoprotein

“…Six MAbs (IgG1Î), designated AFY1-AFY6 (Y1-Y6), were described previously [6]. Thirty purified MAbs, ISOBM 92-121, were supplied through the ISOBM TD-2 AFP Workshop [8].…”

“…Using the same workshop panel of 30 MAbs in addition to 6 MAbs from our own laboratory [6], we character- The primers were designed to create a BglII site (AGATCT) and a SacI site (GAGCTC) at the end of the products.…”

“…Since hepatoma and yolk sac tumors produce this protein frequently and specifically, this protein has been routinely used as an excellent tumor marker for diagnosis as well as for basic studies of malignancies [1,2]. Polyclonal and monoclonal antibodies (MAbs) against human AFP have been used in a variety of procedures such as immunoassay, affinity chromatography, immunoblotting, immunohistology, immunoscintigraphy and immunotherapy [3][4][5][6][7]. In some studies, MAbs were shown to have several advantages over polyclonal antibodies.…”