High-sensitivity detection of autoantibodies against proteinase-3 by a novel third-generation enzyme-linked immunosorbent assay.

The aim of this study was to evaluate a novel third-generation enzyme-linked immunosorbent assay (ELISA) for the high-sensitivity detection of autoantibodies to proteinase-3 (PR3) in patients with Wegener's granulomatosis (WG). First- and second-generation ELISA for the detection of antineutrophil cytoplasmic antibodies (ANCA) frequently demonstrate insufficient sensitivity due to inadequate presentation of autoantigenic epitopes. Human PR3 was immobilized on the solid phase of ELISA plates by anchoring technique.

Serological differentiation of inflammatory bowel diseases.

Objective: In order to shorten the diagnostic procedures simple serological tests with high diagnostic value for Crohn's disease and ulcerative colitis are required. The aim of this study was therefore to evaluate the diagnostic value of a newly developed assay for the determination of antibodies to mannan of Saccharomyces cerevisiae in combination with antibodies to antigens of exocrine pancreas, goblet cells, human neutrophils and tissue transglutaminase.

Methods: Sera from 112 patients with Crohn's disease (82 confirmed and 30 suspected cases), 65 patients with ulcerative colitis, 13 patients with indeterminate colitis, 212 patients with other diseases and 250 healthy blood donors were tested for IgA and IgG antibodies to mannan of S.

Monoclonal antibodies to human chorionic gonadotropin (HCG) and their use in two-site binding enzyme immunoassays.

A panel of mouse monoclonal antibodies (MABs) was produced against human chorionic gonadotropin (HCG) and its isolated beta-subunit (beta-HCG). According to their binding specificities the antibodies could be divided into HCG-specific and cross-reactive MABs. The HCG-specific antibodies reacted with antigenic sites on holo-HCG or holo-HCG and beta-HCG, or exclusively with the non-associated beta-HCG chain.

[Tetramethylbenzidine--a chromogenic substrate for peroxidase in enzyme immunoassay].

3,3' 5,5'-tetramethylbenzidine (TMB) is a high sensitive chromogenic substrate for horseradish-peroxidase as a marker enzyme. In an enzyme immunoassay (EIA) for alpha-1-fetoprotein and in an rapid EIA for myoglobin it reveals higher sensitivity compared to o-phenylenediamine, the increase depends on reaction time. The optimal peroxide concentration depends on reaction time of enzyme chosen in different assays.

Development of an immunoenzymometric assay for alpha 1-microglobulin and measurement of its serum concentration in normal and HIV-infected persons.

alpha 1-Microglobulin was purified from urine to a purity of 97.7% in a yield of 25.8%, and was used to produce antibodies in sheep.