Composition and method for detecting malignant neoplastic disease
A composition and complex technology, applied in chemical instruments and methods, disease diagnosis, biomaterial analysis, etc., can solve the problems of lack of organ specificity, elevated serum levels, etc.
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Embodiment 1
[0381] Assay for the Determination of Heterocomplexes of Keratin 7 and Keratin 19 in Body Fluids
[0382] The aim is to generate an assay suitable for clinical evaluation.
[0383] According to the method described in "Recent development in the periodate method of conjugating horseradish peroxidase (HRP) to antibodies" (M Barbara Wilson and Paul K. Nakane, Elsevier / North-Holland Biomedical Press, 1978), the keratin 7-specific The monoclonal antibody Ks 7.18 was conjugated to biotin and the monoclonal antibody BM 19.21 specific for keratin 19 was conjugated to peroxidase.
[0384] A sandwich enzyme-immunoassay was performed to determine the heterocomplexity of keratin 7 with keratin 19 in body fluids obtained from patients diagnosed with a malignant neoplastic disease or from patients diagnosed with a non-malignant disease or individuals considered healthy substance or fragments thereof. In this procedure, two monoclonal antibodies (biotin-conjugated Ks 7.18 (1 μg / ml) and per...
Embodiment 2
[0386] Sensitivity of the K7 / K19 assay for different forms of cancer
[0387] A total of 398 sera from patients with various forms of cancer were tested using the K7 / 19 assay described in Example 1. The results are summarized in Table 1.
[0388] In conclusion, the K7 / K19 assay showed the highest clinical sensitivity for bladder, ovarian, lung, and esophageal cancers compared to other cancers studied, and especially samples from lung, bladder, and ovarian cancer patients had high concentrations of keratin Heterocomplex of 7 with keratin 19 (or a fragment thereof). In contrast, the K7 / K19 assay showed sensitivities as low as 10% or less for squamous cell and breast cancers of the cervix, as well as head and neck and gastrointestinal cancers. The concentration values of the heterocomplex of keratin 7 and keratin 19 (or fragments thereof) of these latter cancer patients were in the same range as those of the controls (healthy and benign) in Table 2. K7 / K19 assays are therefo...
Embodiment 3
[0392] Establishment of reference ranges for healthy individuals.
[0393] One hundred and twelve (112) sera from apparently healthy individuals were tested according to the K7 / K19 assay as described in Example 1 in order to establish a reference range for individuals with no overt signs of malignant neoplastic disease. 95% of the individuals had a concentration of the heterocomplex of keratin 7 and keratin 19 or fragments thereof of less than 1.01 U / mL, or 95.5% had a value of 1.09 U / mL or less. Five (5) of 112 healthy individuals (ie 4.5%) had concentrations above 1.1 U / mL. The average concentration of the heterocomplex of keratin 7 and keratin 19 or fragments thereof in the test individuals was 0.15 U / mL (see Table 2).
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