A New Immunoassay for Quantification of a Novel Cancer Antigen in Serum and Immunostaining of Carcinoma Tissues and Cultured Cells Revealing the Antigenic Cellular Location.
The purpose of this study is 1) to examine the presence of the antigen in serum by employing a newly developed ELISA immunoassay that quantifies the total antigen and bound antigen (antigen-antibody complex) using polyclonal chicken antibodies directed against an IgM-binding epitope of the new antig...
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| Format: | Others |
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Digital Commons @ East Tennessee State University
2002
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| Online Access: | https://dc.etsu.edu/etd/719 https://dc.etsu.edu/cgi/viewcontent.cgi?article=1876&context=etd |
| Summary: | The purpose of this study is 1) to examine the presence of the antigen in serum by employing a newly developed ELISA immunoassay that quantifies the total antigen and bound antigen (antigen-antibody complex) using polyclonal chicken antibodies directed against an IgM-binding epitope of the new antigen, and 2) to determine the location of the antigen in carcinoma and normal cells. Sera from healthy volunteers (n = 147) and cancer patients (n = 26) were compared for both bound and total antigen concentrations using the new ELISA. Healthy volunteers were subdivided into three groups: those with a personal history of cancer (n = 13), those with no personal or family history of cancer (n = 36) and those with a family history of cancer (n = 97). Ovarian, breast, colon carcinoma tissues and their normal counterparts and cultured ovarian and prostatic carcinoma cells were subjected to immunofluorescence using IgY antibodies and goat anti-chicken fluorescent secondary antibodies. Basic imaging was performed on tissue sections while confocal microscopy was performed on cultured cells. Furthermore, immunohisto-chemical staining using an anti-chicken HRP-conjugated secondary antibody was performed on 16 normal ovarian tissues, 53 ovarian adenocarcinomas, and 3 borderline ovarian tumors. Statistical analysis revealed significant differences in cancer patients' bound and total antigen levels compared to that of healthy volunteers (p < 0.005). Bound and total antigen levels of cancer patients were also significantly higher than those of the healthy volunteers with no personal or family history of cancer and those with a family history of cancer (p < 0.01). However, no significant difference existed between the bound (p > 0.120) and total antigen levels (p > 0.076) of cancer patients and patients with a personal cancer history. Immunohistochemical staining of ovarian tissues revealed a significant difference in the lumenal staining of the carcinomas compared to that of the normal ovarian tissues. Furthermore, fluorescence imaging revealed that the antigen is localized to the cell membranes of the carcinoma cells but is absent from the normal tissues. Confocal microscopy further emphasized the antigen's association with the membrane and also revealed some filamentous cortical staining. |
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