[en] In 1975 Bartos et al described a radio-immunoassay for spermine which lacked specificity. In subsequent years the same group has reported the production of a specific antibody to spermidine and to spermine. The availability of specific antibodies potentially transformed the usefulness of radioimmunoassay from a system useful for semi-quantitative screening to one with a much broader applicability in the area of quantitative assay of polyamines. It was claimed that the assay would quantify as little as 10 picogrammes of spermidine or spermine. Up to 1982 when the present project commenced no other groups had reported using the Bartos assay. The present study was therefore indicated in order to determine whether specific antibodies could be produced in laboratories other than those of the Bartos group and to validate the assay by comparing it with the much more widely used and well established HPLC methods currently available

[en] A solid phase radioimmunoassay has been developed which can be used for the detection of isotype specific antibodies to whole bacteria and other particulate antigens, and is applicable to a variety of species. Bacteria are bound to the solid phase by the use either of antibodies, or of methyl glyoxal. Both methods result in a sensitive and reproducible assay, and bacteria do not appear to desorb from the solid phase. The specificity of antibodies to whole bacteria was examined by absorption of antisera with various species of bacteria and retesting, or by determining the binding of antisera to various bacteria bound to the solid phase. Both methods revealed specificity for the bacteria examined. Inhibition studies showed that antibodies to Streptococcus mutans whole cells could be inhibited by purified cell surface antigens glucosyltransferase and antigen I/II, but only minimally by lipoteichoic acid, c polysaccharide or dextran. In murine antisera antibodies of the IgG, IgM, and IgA classes could be detected at amounts of less than 1 ng/ml. (author)

No abstract available

[en] Monoclonal antibodies (MAbs) reactive with Marburg virus (strain Musoke) were evaluated for both biological activity and specificity. Several of the Marburg virus- (MBGV) specific MAbs reduced the size and/or number of MBGV plaques in vitro. The ability of the MAbs to affect plaque formation in vitro was demonstrated to be specific for the glycoprotein (GP) of the strain of MBGV used for vaccination. Using deletion analysis and peptide mapping, the binding epitopes of several of these neutralizing MAbs were identified. Not unexpectedly, the epitopes were shown to lie in the most hypervariable and highly glycosylated region of MBGV GP. An analysis of the in vivo activity of several MAbs revealed that some antibodies provided substantial but incomplete protection of naive guinea pigs by passive transfer. These data suggest that neutralizing epitopes exist within MBGV GP but that induction of antibodies to these neutralizing epitopes may not be sufficient for protection from lethal infection

[en] A unitized, solid phase kit for radioimmunoassay is described. All of the necessary assay reagents are incorporated in a single tube. Requiring only the addition of the patient's sample, all phases of the assay procedure are performed in this tube. Antibodies are bound to the tube surface, while labelled antigens are also present but unbound. Storage in the absence of air and water results in the stabilization of the reagents such that the system can be stored for long periods

[en] The aims of this study were to generate chimeric human papillomavirus (HPV)-16 L1 virus-like particles (VLPs) in order to identify immunogenic domains and conformational neutralizing epitopes, and to characterize the regions where a foreign epitope could be introduced. We hypothesized that these regions could be on L1 protein loops since they are exposed on the surface of VLPs. The aims of this study were achieved by mutating HPV-16 L1 proteins. Six amino acids encoding for the epitope 78-83 (DPASRE) of the hepatitis B core (HBc) antigen were introduced within the different loops of the L1 protein at positions 56/57, 140/141, 179/180, 266/267, 283/284 or 352/353. All these chimeric L1 proteins were capable of self-assembly into VLPs. The antigenicity and immunogenicity of some of these VLPs were reduced compared to the levels observed with wild-type VLPs. All were nevertheless able to induce neutralizing antibodies. VLPs with insertion at position 266/267 induced lower levels of neutralizing antibodies, suggesting the involvement of residues situated on FG loop in L1 neutralizing epitopes. All the chimeric L1 proteins except the one with insertion at position 56/57 were also able to induce anti-HBc antibodies, thus suggesting exposure of the HBc epitope on the VLP surface. Taken together, our findings indicate the possibility of designing HPV-derived vectors that are less immunogenic and suggest positions for insertion of defined immune epitopes or cell ligands into L1 protein to be exposed on the surface of VLPs

[en] Previous studies from our laboratory have demonstrated the feasibility of immunoassays for identification and quantification of specific metal ions. Our ultimate goal for this project is to (1) isolate and characterize antibodies that recognize the most mobile form of uranium, UO22+; (2) assemble, test, and validate a new field-portable immunosensor based on these antibodies; (3) prepare new monoclonal antibodies to the primary chelators (EDTA and DTPA) found in DOE wastes

No abstract available

[en] A rapid multisite immunoradiometric assay for measurement of human α-fetoprotein (AFP) that uses a high affinity monoclonal and polyclonal antibodies directed against distinct and separate determinants on the protein was developed and designated as coated-tubes IRMA (CT-IRMA). AFP standards or serum samples were added into the polystyrene tubes coated with goat IgG antibody against human AFP. After 1 hour incubation, the tubes were washed and a radioiodinated mouse monoclonal antibody was added and then incubated overnight at room temperature. The sensitivity of the CT-IRMA was found to he 0.26 ng/ml. The recovery of AFP mixed with human serum was 98.1 -111.1 %. The within-assay, and between-assay coefficients of variation were 1.49 - 3.11 % and 4.89 - 7.44 %, respectively. The assay correlated well with a commercial method (CIS bio international, France) ( r = 0.9741, n = 83, OAEP kit 0.8748 CIS kit - 2.1561). The mean concentration of AFP in normal serum was 5 ng/ml. This developed kit reagents are being assessed clinically in multicenter study at National Cancer Institute, Chulalongkorn Hospital and Pramongkutklao Hospital to find the real clinical application in various groups of patients which include early detection monitoring therapy, follow up of hepatocellular carcinoma, early identification and monitoring of AFP- producing tumors in high-risk populations. (Author)

[en] Two monoclonal antibodies were raised against human gamma interferon (IFN-γ) derived from E. coli harboring the recombinant cDNA for IFN-γ, and one against a synthetic peptide representing its C-terminus amino acid sequence of 20 residues. The monoclonal antibody against the synthetic peptide reacted either with IFN-γ or the synthetic peptide. One monoclonal anti-IFN-γ did not react with the synthetic peptide, while the other showed a weak binding with the peptide. A 2-site '1-step' radioimmunoassay was developed. The assay was rapid with a sensitivity capable of detecting a few ng/ml of IFN-γ. (Auth.)