“…24 of 46 (52%) ovarian cancer patients, 7 of 18 (39%) patients with benign ovarian cysts or tumors and 3 of 85 (4%) control samples had elevated serum NB/70K levels. Comparison of NB/-70K levels measured in the NB12123 assay with levels measured in an assay using a polyclonal antiNB/70K previously developed in our laboratory [13] indicated that although both assays had approximately the same percentage of positive ovarian cancer patient samples, there appeared to be no correlation between the absolute NB/70K levels measured by the two assays. The rank of ovarian cancer patient samples was also different for the two assays.…”
mentioning
confidence: 76%
“…concanavalin A-Sepharose 4B (Pharmacia, Uppsala, Sweden) [13]. Bound antigen was eluted with a solution of 10% D-mannose in 0.1 M acetate, pH 6.3, containing 1 M sodium chloride, 1 mM CaCI2, McC12 and MgCI 2 and 0.01% merthiolate.…”
Section: Introductionmentioning
confidence: 99%
“…Antigen for use in radioimmunoassays was prepared as follows: antigenic fraction OCC, partially purified NB/-70K, was prepared as previously described [13] and used as labeled antigen for the testing of supernatants from the hybridoma cultures. In addition, NPIF was prepared from outdated blood bank plasma by precipitation with 50% saturated ammonium sulfate, precipitation with perchloric acid, and chromatography on concanavalin A-Sepharose 4B, and used as labeled antigen for the selection of hybridomas which would not bind to NPIF.…”
Section: Introductionmentioning
confidence: 99%
“…A rabbit antiNB/70K was prepared [13] and a radioimmunoassay for detecting serum NB/70K, the triton NB/70K assay, was developed [12]. This assay was used in preliminary experiments to assess the specificity of NB/70K expression in malignant ovarian tumors [10].…”
Murine monoclonal antibodies (MCAs) against human ovarian tumor associated antigen NB/70K have been prepared. One of these MCAs, NB12123, was chosen for the development of a radioimmunoassay for measuring serum NB/70K levels. In this assay, the average NB/70K level in 75 normal, healthy controls was 11.9 activity units (AU) with an SD of 14.9 AU. The normal cut off value for this assay was set at 45 AU (mean +2 SD). 24 of 46 (52%) ovarian cancer patients, 7 of 18 (39%) patients with benign ovarian cysts or tumors and 3 of 85 (4%) control samples had elevated serum NB/70K levels. Comparison of NB/70K levels measured in the NB12123 assay with levels measured in an assay using a polyclonal antiNB/70K previously developed in our laboratory [13] indicated that although both assays had approximately the same percentage of positive ovarian cancer patient samples, there appeared to be no correlation between the absolute NB/70K levels measured by the two assays. The rank of ovarian cancer patient samples was also different for the two assays. Also, almost 40% of patients with benign ovarian cysts and tumors had elevated serum NB/70K levels as measured by the NB12123 assay as compared to 0% for the polyclonal assay. Reciprocal cross-blocking experiments, absorption studies, and immune precipitate analysis indicated that both the monoclonal NB12123 assay and the polyclonal antiNB/70K assay measured the same population of NB/70K molecules. However, the polyclonal antibody recognizes epitopes in addition to that recognized by NB12123. Taken together, these results suggest that the epitope recognized by NB12123 is not as specific for malignant ovarian tumors as the epitope(s) recognized by polyclonal antiNB/70K and/or that more than the one epitope detected by the MCA is responsible for the specificity for ovarian cancer of the polyclonal NB/70K assay. In spite of this, the greater sensitivity and range of the monoclonal NB12123 assay make it possible to monitor serum NB/70K levels in ovarian cancer patients. In four patients examined, the fluctuating serum NB/70K levels appeared to correlate well with clinical status.
“…24 of 46 (52%) ovarian cancer patients, 7 of 18 (39%) patients with benign ovarian cysts or tumors and 3 of 85 (4%) control samples had elevated serum NB/70K levels. Comparison of NB/-70K levels measured in the NB12123 assay with levels measured in an assay using a polyclonal antiNB/70K previously developed in our laboratory [13] indicated that although both assays had approximately the same percentage of positive ovarian cancer patient samples, there appeared to be no correlation between the absolute NB/70K levels measured by the two assays. The rank of ovarian cancer patient samples was also different for the two assays.…”
mentioning
confidence: 76%
“…concanavalin A-Sepharose 4B (Pharmacia, Uppsala, Sweden) [13]. Bound antigen was eluted with a solution of 10% D-mannose in 0.1 M acetate, pH 6.3, containing 1 M sodium chloride, 1 mM CaCI2, McC12 and MgCI 2 and 0.01% merthiolate.…”
Section: Introductionmentioning
confidence: 99%
“…Antigen for use in radioimmunoassays was prepared as follows: antigenic fraction OCC, partially purified NB/-70K, was prepared as previously described [13] and used as labeled antigen for the testing of supernatants from the hybridoma cultures. In addition, NPIF was prepared from outdated blood bank plasma by precipitation with 50% saturated ammonium sulfate, precipitation with perchloric acid, and chromatography on concanavalin A-Sepharose 4B, and used as labeled antigen for the selection of hybridomas which would not bind to NPIF.…”
Section: Introductionmentioning
confidence: 99%
“…A rabbit antiNB/70K was prepared [13] and a radioimmunoassay for detecting serum NB/70K, the triton NB/70K assay, was developed [12]. This assay was used in preliminary experiments to assess the specificity of NB/70K expression in malignant ovarian tumors [10].…”
Murine monoclonal antibodies (MCAs) against human ovarian tumor associated antigen NB/70K have been prepared. One of these MCAs, NB12123, was chosen for the development of a radioimmunoassay for measuring serum NB/70K levels. In this assay, the average NB/70K level in 75 normal, healthy controls was 11.9 activity units (AU) with an SD of 14.9 AU. The normal cut off value for this assay was set at 45 AU (mean +2 SD). 24 of 46 (52%) ovarian cancer patients, 7 of 18 (39%) patients with benign ovarian cysts or tumors and 3 of 85 (4%) control samples had elevated serum NB/70K levels. Comparison of NB/70K levels measured in the NB12123 assay with levels measured in an assay using a polyclonal antiNB/70K previously developed in our laboratory [13] indicated that although both assays had approximately the same percentage of positive ovarian cancer patient samples, there appeared to be no correlation between the absolute NB/70K levels measured by the two assays. The rank of ovarian cancer patient samples was also different for the two assays. Also, almost 40% of patients with benign ovarian cysts and tumors had elevated serum NB/70K levels as measured by the NB12123 assay as compared to 0% for the polyclonal assay. Reciprocal cross-blocking experiments, absorption studies, and immune precipitate analysis indicated that both the monoclonal NB12123 assay and the polyclonal antiNB/70K assay measured the same population of NB/70K molecules. However, the polyclonal antibody recognizes epitopes in addition to that recognized by NB12123. Taken together, these results suggest that the epitope recognized by NB12123 is not as specific for malignant ovarian tumors as the epitope(s) recognized by polyclonal antiNB/70K and/or that more than the one epitope detected by the MCA is responsible for the specificity for ovarian cancer of the polyclonal NB/70K assay. In spite of this, the greater sensitivity and range of the monoclonal NB12123 assay make it possible to monitor serum NB/70K levels in ovarian cancer patients. In four patients examined, the fluctuating serum NB/70K levels appeared to correlate well with clinical status.
“…Carcinoembryonic antigen is a gly coprotein with a molecular weight of 180 kDa [19], However, a number of other markers that might be associated with ovarian carci noma have been described. These include DF-3 [20], CA 15-3 [21], TAG 72 [22], HMFG [23], NB/70K [24] and others. Unfor tunately, we have no information concerning their expression pattern in tissues and molec ular characteristics.…”
A new monoclonal antibody (MAb 9H8, IgM class) reactive with human ovarian carcinoma has been raised after immunizing C57BL/6 mice with bovine sperm. Immunohistological studies indicated that 20/21 serous ovarian adenocarcinomas expressed 9H8-defined antigen but it was absent in benign ovarian tumors (0/11). 1/11 of breast carcinomas and 5/5 of rectal carcinomas expressed this antigen, although to a considerably lesser degree. Tumors of lung, skin, brain and mesothelium were negative. The antigen was also expressed in embryonic skin, in renal collecting tubule cells and in saliva. In bovine, human and mouse sperm the antigen is confined to the acrosomal region. The molecular weight of this antigen was determined by Western blot analysis and gel filtration. In SDS-PAGE the antigen ran as a broad band barely entering the 7% gel, indicating an apparent molecular weight > 300 kDa. In the absence of detergents and reducing agents this glycoprotein forms larger complexes ( > 1,500 kDa) as determined by gel filtration on Sephacryl S300. The epitope contains carbohydrate structures recognized by lectin PNA (peanut agglutinin).
Blood levels of human ovarian tumor antigen NB/70K were measured using four different monoclonal anti-NB/70K antibody radioimmunoassays (NB12123, NB12913, NB13834, and NB13831 assays). Four hundred thirty-two blood samples were obtained from apparently disease-free controls, from patients with benign and malignant ovarian tumors, and from patients with benign nontumorous and malignant diseases of the breast, lung, and gastrointestinal systems. The means, medians, and percent of samples that had elevated levels of NB/70K were calculated for each group of patients or controls for each assay. For all control versus malignant disease comparisons, there were statistically significant differences in all assays. For all benign versus malignant disease comparisons in the ovary, breast, lung, and gastrointestinal systems, there were statistically significant differences in all assays with the exception of the benign versus malignant gastrointestinal disease comparison in the NB12123 assay. Elevated NB/70K levels were detected in blood from ovarian cancer patients, almost all of whom had adenocarcinomas. Elevated NB/70K levels were also detected in blood samples from patients with adenocarcinoma, squamous, and clear cell carcinomas of the lung as well as in adenocarcinoma and ductal carcinoma of the breast. These results indicate that NB/70K is a marker for different pathologic types of malignancies.
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