Identification of Isoform p-53 Protein in CLL by Elisa Method

The monoclonal p-53 antibody, PAb 240, used in the ELISA method recognizes both mutant and wild-type p-53 under denaturing conditions. Species reactivity is for human or rhesus monkey in conformity with the prospect. The monoclonal antibody PAb 240 recognizes an epitope that is structurally hidden in the wild-type conformation of p-53 and becomes exposed by denaturing the p-53 protein or the mutant conformations of p-53, where point mutations in the P-53 gene alter the terminal structure of the p-53 protein. This ELISA kit is recommended for use in serum, plasma or tissue homogenates. Using other types of sample is not supported. The sample collection protocols below were adapted from the references. Results Of the 20 patients studied, 14 men have aged 55-85 years and 6 women aged 39-85 years. Patients were treated at the time of these investigations with cytostatic and immunotherapy specific for CLL. For the men the results of protein concentration p-53 detection were following values, 20, 15, 18, 40, 10, 12, 14, 60, 30, 10, 13, 13, 5, 10, 15, 12, (µg / dl) and women's results of protein concentration p-53 were 140, 30, 13, 10, (µg / dl), The pathological values in the 3 cases of highly elevated p-53, reflecting the concentration of p-53 protein mutant, were calculated in 2 men in the amount of 60 µg / dl, respectively in 50 µg / dl, and in the female case, it was calculated in the amount of 140 µg / dl, the frequency of chronic lymphocytic leukemia being higher in men than in women, in ratio 2/1, in accordance with the data from the specialized literature. The overall frequency of positivity of the p53 protein, in the increased number of CLL cases studied, was 15% (3 out of 20 cases). The expression of the high-concentration p53 proteins in stage 2/3 of the disease was associated with a significantly weaker response to chemotherapy (p = 0.034). Statistical interpretations: the concentration of p-53 protein, in the 17 cases with p-53 protein expression, after excluding the 3 out-line cases present in the study, was calculated at the average value of 16.76µg / dL. In the CLL stage III/IV (n = 3 patients (Table 1), In the presented work with CLL, in the stage I/II, (n=17 patients) was registered in the average p-53 protein concentration in CLL, 16.76, (µg/dl), with mean values of hemogram: hematological parameters in peripheral blood, Leukocytes numberer = 35-50 x 10 3 / dL, Hb = 11.8g /dL; Platelet = 140 x 103/dL, Lymphocytes in peripheral blood = 65-80%. In some international studies, the immunological characteristics of patients with CLL having p-53 protein positive, were measured by immunohistochemistry, (IHC). In the second part of the study CLL on the stage I/II, (n=47 patients), the P-53 protein isoform concentration in reactive Lymphocyte B, the average p-53 protein concentration in CLL was on average 47 U/m with 16.7% samples studied, (7 out of 42 cases).