ABSTRACT
This paper is the first report on the use of an idiotype-anti-idiotype monoclonal antibody reaction to develop an enzyme immunoassay for thyroxine (T4). We have developed a monoclonal antibody against T4, named 1F10 of IgG1 subclass and KA 5.21 x 10(8) M-1 which was used to obtain anti-idiotypic monoclonal antibodies. Anti-idiotypic antibodies were selected by a novel method, a passive agglutination assay with the idiotype monoclonal 1F10 absorbed on latex particles and subsequently characterized by RIA. One of these anti-idiotype antibodies, named 5B3--type beta antibody--of IgG1 subclass, was used to develop an enzyme-linked T4 idiotype-anti-idiotype immunosorbent assay. The T4 calibration curve, using the 1F10 idiotypic antibody adsorbed to solid phase and the 5B3 anti-idiotypic antibody conjugated to alkaline phosphatase (ALP), shows adequate performance in the range between 0.7-25 micrograms% of the analyte. The reliability of the proposed method is demonstrated by the correlation coefficient r = 0.74, found between T4 measured by RIA and our assay, with a panel of sera from euthyroid, hypothyroid and hyperthyroid individuals. The correlation coefficient was r = 0.93 within assays and r = 0.88 between assays. These results provide the basis for a new non isotopic assay for the study and diagnosis of T4-related human disease and provides a model to develop immunoassays for other haptens and small molecules of clinical interest.
Subject(s)
Antibodies, Anti-Idiotypic/chemistry , Antibodies, Monoclonal/chemistry , Enzyme-Linked Immunosorbent Assay/methods , Immunoglobulin Idiotypes/chemistry , Thyroxine/blood , Thyroxine/immunology , Animals , Antibody Affinity , Female , Mice , Mice, Inbred BALB CABSTRACT
An improved procedure for the generation of high-avidity anti-human B blood group monoclonal antibodies (MAbs) was developed. One of them, termed 7A1-2, showed excellent qualities of titer, avidity, and intensity required for use as human B blood typing reagent. Hemagglutination inhibition studies with monosaccharides and oligosaccharides were carried out to determine the specificity of the MAb 7A1-2. These studies indicate that the antibody reacts with the immunodominant region of the antigen which is known to confer the serologic specificity of this blood group.
Subject(s)
ABO Blood-Group System/immunology , Antibodies, Monoclonal/immunology , Blood Grouping and Crossmatching/methods , Isoantibodies/immunology , Animals , Antibody Affinity/immunology , Antibody Specificity/immunology , Female , Hemagglutination Tests , Humans , Indicators and Reagents , Mice , Mice, Inbred BALB CABSTRACT
Industrial processes may lead to severe alterations of environment affecting the health and quality of life of neighbouring communities. The present study reports the concentrations of Cu, As, Br, Na, K, Se, Hg, Cr, Co, Zn, Fe and Sb in samples of hair from children 6 years of age living in Antofagasta, Chuquicamata, María Elena, Calama and San Pedro de Atacama. Results were compared to those obtained in children of the same age living in Santiago. Samples were obtained according to recommendations from the International Nuclear Energy Organization and were analyzed by the neutron activation method at facilities of the Chilean Nuclear Energy Commission. Values for Cu (38.8 ppm), Cr (4.29 ppm) and As (19.2 ppm) in children from Chuquicamata are considered dangerous according to international standards.
