XXXX @entity71080 : production and purification of recombinant protein and monoclonal and polyclonal antibodies, and development of a sensitive and specific immunofluorometric assay.

@entity1 @entity825 @entity43 @entity71080 @entity10056 @entity7435 @entity375
BACKGROUND: The aims of this study were to develop immunologic reagents and a sensitive and specific immunoassay for @entity1 @entity71080 ( @entity7435 ) and to examine the presence of @entity7435 in @entity1 tissues and biological fluids. METHODS: Recombinant @entity7435 protein was produced and purified with use of a @entity10056 @entity375 expression system. The protein was used as an immunogen to generate mouse monoclonal and rabbit polyclonal anti- @entity7435 antibodies. A sandwich-type immunoassay was developed with these antibodies. The assay was used to measure @entity7435 in various biological fluids and tissue extracts. Immunohistochemical analysis was also performed on nondiseased and cancerous prostatic sections. RESULTS: The @entity7435 immunoassay had a detection limit of 0.05 micro g/L and showed no cross-reactivity with homologous kallikreins. The assay was linear at 0-20 micro g/L, and within-and between-run CVs were <10% (n = 12). @entity7435 was detected in tissues, including esophagus, tonsil, @entity825 , lung, cervix, and prostate. @entity7435 was also found in seminal plasma, amniotic fluid, follicular fluid, ascites of @entity43 @entity1 , breast milk, and cytosolic extracts of @entity43 tissues. @entity7435 was immunohistochemically localized in epithelial cells of both nondiseased and cancerous prostate. @entity7435 appears to be overexpressed in 50% of @entity43 tissues compared with healthy ovarian tissues. Recovery of active enzyme added to milk or amniotic fluid was 70-98%, but was <20% when added to serum, suggesting rapid sequestration by protease inhibitors. In fluids and tissue extracts, @entity7435 was found in its free (approximately 30 kDa) form. CONCLUSIONS: This immunofluorometric assay for @entity7435 may be used to examine the value of @entity7435 as a disease biomarker and to further explore the physiologic and pathobiologic role of this enzyme in @entity1 disease.
