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Biomedical subjects

F Celada

Publications and source records attributed to F Celada.

At least 73 records · Page 4Linked to original sources
PubMed1976

An immuno-enzymatic assay of cortisol using E. coli beta-galactosidase as label.

An enzyme-assay of cortisol was established by using E. coli beta-galactosidase as a marker. The number of cortisol residues per molecule of enzyme and the method of separation of free from antibody-bound enzyme proved to be critical factors for the assessment of the assay. A sensitivity ranging from 100 to 150 pg of cortisol per tube has been achieved. A comparsion with other methods, i.e. competitive protein binding assay, radioimmunoassay and fluorimetry, is reported.

Animals↗
PubMed1976

An in vitro immuno-enzymatic assay of tumor antigens in the mouse with beta-galactosidase.

A method for detection of the primary binding of soluble tumor-associated antigens by antibodies has been developed by using an enzyme immunoassay (EIA). A heteroantiserum was produced by injecting tumor cells from a chemically induced murine sarcoma into rabbits, and antibodies reacting with most normal tissue components were removed by exhaustive in vivo absorption. A soluble preparation of tumor cells, obtained by 3 M KCl extraction, was conjugated to beta-galactosidase from Escherichia coli. The antibody binding was measured by determining the enzyme activity that could be separated by anti-antibody coprecepitation. The reaction follows saturation kinetics, and nonlabeled antigen can be readily quantitated by inhibition. The present method detects determinants common to several MC-induced tumors on the same mouse strain but absent in normal cells and nonrelated tumors in addition to individual tumor-specific transplantation antigens. The sensitivity and simplicity of the new method compare favorably with a binding assay that utilizes radioactive iodine as a label. Thus, EIA becomes a flexible tool for the further characterization and purification of these antigens.

Animals↗
PubMed1972

Maturation of the immkune response in vitro. Focal fluctuation and changes in affinity of anti-beta-D-galactosidase activating antibody.

We have cultivated lymph node microfragments from beta-D-galactosidase (Escherichia coli) primed rabbits and have measured their secondary response directed towards the whole molecule (precipitating antibodies) and to a single determinant (activating antibodies) of the antigen. By decreasing the size of the fragments to 10(5) cells, we began to observe heterogeneity among identical cultures in terms of positivity of response, antibody specificity, and titers. The affinity of "early" activating antibodies was inversely proportional to the dose of challenge. While no maturation was seen in low and excessive challenge, in all cultures receiving intermediate doses the association constant was raised several orders of magnitude within periods of 20 days. The relevance of these data to the mechanism of affinity selection of antigen-sensitive cells is discussed.

Animals↗
PubMed1971

Antibody-mediated activation of a defective beta-D-galactosidase. II. Immunological relationship between the normal and the defective enzyme.

Two closely related protein antigens were used to study immunogenic competition. Namely, normal beta-D-galactosidase of Escherichia coli (Z) and a genetically defective beta-D-galactosidase (AMEF) which seems to differ from the normal in one amino acid substitution. A unique characteristic of this pair of antigens is that, although they are indistinguishable in precipitation and absorption tests with antibodies, the enzymatic activity of AMEF is specifically increased several-hundredfold in the presence of antibodies directed against Z. The following results show that Z and AMEF also differ in their immunogenic ability: (a) antibodies directed against Z activated AMEF; antibodies directed against AMEF did not activate, but competed specifically with activating antibodies. (b) Animals immunized with AMEF failed to produce activating antibodies when they were subsequently challenged with Z, although the presence of some cells primed to produce activating antibodies could be demonstrated by adoptive transfer. (c) Animals preimmunized with Z were stimulated in their production of activating antibodies by AMEF challenge, although not as efficiently as with Z. A model explaining these observations by competition for the immunogenic site among antigen-sensitive cells carrying cross-reacting receptors is presented.

Adoptive Transfer↗