Tumor-associated protein survivin is the bifunctional protein which can participate either in cell division regulation or in apoptosis inhibition depending on its localization and structure state. The aim of this work was to obtain monospecific antibodies useful for investigation of protein structure and functional features. Six affinity purified antibodies directed to different protein regions were obtained. The ability of antibodies obtained to detect survivin in tumor cells and breast cancer tissues was studied. It was shown that antibodies to (1-22) and (95-105) survivin fragments have the highest specific activity. In western-blot antibodies to (1-22) region predominantly binds with survivin-containing complex, which may be the survivin dimer as we suppose, while antibodies to (95-105) region detects only monomeric form of the protein. Breast cancer tissues study demonstrated that survivin monomer presents only in the tumor core tissues, while survivin-containing complex is expressed both in tumor core and tumor periphery tissues. It was shown that antibodies to (1-22) fragment detect predominantly nuclear survivin, which participates in mitosis regulation, while antibodies to (95-105) fragment gave nucleoplasm and cytoplasm staining at all stages of cell cycle. Thereby antibodies obtained are the useful tool for structure-functional study of survivin.

For today, one of actual problems in gynecologic pathology are borderline tumours of the ovary to which frame of a case rate the high specific gravity is compounded by women of reproductive age. Thereupon often there is a question about safety treatment methods, that dictates necessity of an assessment of risk of development of recurrence of disease. In present article attempt to define prognostic importance of markers of proliferative activity (ki67, cyclin D1, p21), and pro-and antiapoptotic markers (p53, bax, bcl-2, survivin) for definition potentially an aggressive behavior of ovarian borderline tumour.

Tumor-associated protein survivin is a bifunctional protein that can participate either in the regulation of cell division or in the inhibition of apoptosis depending on its location in the cell and its structural state. The aim of the present work was to obtain monospecific antibodies to survivin targeting various sites in the protein and suitable for the study of its structural and functional features. The ability of the antibodies to detect survivin in tumor cells and mammary tumor tissues has been assessed. Antibodies targeting the (1–22) and (95–105) fragments of the protein have been shown to have the highest specific activity. Antibodies targeting the (1–22) site preferentially bound to survivin-containing complex (presumably a dimer of survivin) in immunoblotting, whereas antibodies targeting the (95–105) site detected only the monomeric form of the protein. Analysis of malignant mammary tumor samples showed that monomeric survivin was present only in tumor tissue samples, whereas survivin-containing complex was expressed both in tumor tissues and in the tissues adjacent to the tumor. Antibodies targeting the (1–22) site were shown to preferentially detect survivin localized in the cell nuclei (and involved in the regulation of mitosis) in immunocytochemistry, while those targeting the (95–105) site stained the nucleoplasm and cytoplasm at all stages of the cell cycle. Therefore, the antibodies obtained can serve as a useful tool for structural and functional research on survivin.

Whether the expression of the apoptosis inhibitor survivin was correlated with the degree of differentiation and the stage of transitional cell carcinoma of the urinary bladder was studied. Sixty samples of surgical specimens from patients with urothelial carcinomas of various degrees of differentiation and different stages were examined. An immunohistochemical study using the monoclonal antibodies obtained by the authors was conducted. The high expression of survivin was shown to be correlated with the lower-grade differentiation of a tumor and its higher stage.

Survivin, an endogenous protein, is a promising marker for the cancer diagnosis. The aim of the present work was to obtain antibodies specific to survivin and capable of detecting this protein in tumor tissues. Four peptides corresponding to fragments (1–22), (54–74), (80–88)–(153–165), and (118–144) of the survivin-2B sequence were selected and synthesized. Rabbits were immunized with the synthetic peptides. It has been shown that all peptides in a free state, without conjugation with a high-molecular-weight carrier, stimulate the production of antibodies capable of binding with recombinant survivin. Antipeptide antibodies were isolated from sera and their performance in the immunohistochemical detection of survivin in human tumor tissues was studied. It was shown that only antibodies to the (80–88)–(153–165) peptide bind to the survivin present in breast and bladder tumors. The ability of antibodies to this peptide to detect survivin in tumor tissue lysates was demonstrated by immunoblotting. The part of the sequence targeted by the antibodies against the (80–88)–(153–165) peptide was localized using truncated peptide fragments.

Immunoactive fragments corresponding to the N-terminal (19–36) and C-terminal (283–294) regions of the NPM1.1 isoform of nucleophosmin and their shortened fragments were chosen and synthesized. Rabbits were immunized with free full-size peptides and their protein conjugates. Antibodies produced against the 19–36 and 283–294 peptides were purified by affinity chromatography on cyan-bromide activated sepharose that was preliminary conjugated with the synthetic peptides. An analysis of immunoblots of lysates of the HeLa and Ramos cells demonstrated that the antibodies produced against the 19–36 peptide detected the monomeric form of nucleophosmin, whereas the antibodies against the 283–294 peptide predominantly revealed its oligomeric form. It was established by immunocytochemical analysis that the antibodies induced by the 19–36 peptide stained the nucleoplasm and perinuclear space of the cytoplasm of the HeLa and Ramos cells, but did not stain the nucleoli, while the antibodies against the 283–294 peptide stained only the nucleoli of the same cells. On the basis of these results, one could propose that the monomeric and oligomeric forms of nucleophosmin were located in the nucleoplasm and nucleoli of the examined cells, respectively. Thus, antibodies which can predominantly detect monomeric and oligomeric forms of nucleophosmin were produced for the first time. An analysis of the monomeric-oligomeric state and the localization of the nucleophosmin in tumor cells could be performed using these antibodies.