33 results about "Sperm protein" patented technology

The invention discloses a method for screening a biomarker related to severe oligoasthenospermia. The method comprises carrying out deep mass spectrometry on multiple sperm protein uncoded amino acids of severe oligoasthenospermia through a NanoHPLC-MS / MS mass spectrometry system and an unlabeled quantitative proteomics method, searching the mass spectrometry data through a nonfinite amino acid protein modification analysis method, carrying out multivariable gaussian mixture distribution clustering analysis to identify the non-coding amino acids in the sperm protein group as much as possible, and comparing the non-coding amino acids of the normal and patient sperm protein groups to obtain protein uncoded amino acid sites related to severe oligoasthenospermia, wherein the protein uncoded amino acid sites are used as molecular markers of severe oligoasthenospermia. The method provides novel diagnostic and therapeutic targets for severe oligoasthenospermia.

The present invention provides compositions and methods useful for regulating fertilization and for use as a contraceptive based on the discovery herein of an oocyte specific protein, SAS1R (Sperm Acrosomal SLLP1 Receptor), which is a sperm protein receptor. Six SAS1R variants, including the full length SAS1R, were identified. mSLLP1 and SAS1R co-localized to oocytes and to acrosomes of acrosome-reacted sperm. Interactions between mSLLP1 and SAS1R were demonstrated by far-western analysis, in a yeast two-hybrid system under stringent selection conditions, and by immunoprecipitation of SAS1R by anti-mSLLP1 as well as the converse. Purified recombinant SAS1R was found to have protease activity, to inhibit fertilization in-vitro, and to induce an immune response in females. Together, the results suggest SAS1R is a proteolytically active, oocyte and early embryo specific oolemmal metalloprotease receptor for the sperm intra-acrosomal ligand SLLP1 and is a target for regulating fertilization and as a contraceptive.

A 22 kD sperm protein, SP22, correlates with fertility and predicts fertility in males. The protein can be assayed to detect decreases in fertility resulting from exposure to toxicants and pollutants which are known or suspected to decrease fertility. In an antibody is generated to this protein, the antibody recognition by sperm in an epididymal sperm sample or ejaculate would reflect the fertility of the sample. This antibody can be used as a contraceptive to inactivate sperm, screen for toxicity, select animals for artificial insemination, and select men for assisted reproductive technologies. The protein itself can be inactivated by gene knockout, which is another approach to contraception, or the protein can be added to sperm from infertile men to make fertility techniques more feasible.

The invention relates to immunocontraceptive synthetic peptide and immunocontraceptive chimeric peptide, wherein the amino acid sequence of the immunocontraceptive chimeric peptide is shown as SEQ ID:1 in a sequence table, the amino acid sequence of the immunocontraceptive synthetic peptide hESP103-107 is shown as SEQ ID:2 in the sequence table, and the amino acid sequence of the immunocontraceptive synthetic peptide hIzumo216-220 is shown as SEQ ID:3 in the sequence table. The invention also provides the application of the chimeric peptide, the synthetic peptide hESP103-107 or the synthetic peptide hIzumo216-220 as immunocontraceptive vaccines observed in contraception. The invention researches the chimeric peptide comprising sperm protein hESP, hIzumo and tNASP as well as the cross reacting antigens of UU (Ureaplasma Urealyticum) for the first time, and immunized mice can obtains stronger and reversible antifertility effect.

The invention discloses a method for extracting and separating total proteins of mice sperms by using two-dimensional electrophoresis. The method comprises the following steps: preparing sperms, precipitating DNAs of the sperms after lysis of the sperms, preparing sperm proteins, preparing a protein extraction sample, carrying out first-dimensional isoelectric focusing electrophoresis, carrying out second-dimensional SDS-polyacrylamide gel electrophoresis, dyeing by using coomassie brilliant blue, preparing gel and scanning by using an image scanner. According to the method, lysis of the sperms is completely performed by Trizol, the proteins are extracted by chloroform, impurities are removed by isopropanol, guanidine hydrochloride and ethyl alcohol, and the proteins are purified by acetone, so that the problems of low content of the mice sperms, collection difficulty, small cell volume, a small amount of cytoplasm and a large amount of acrosomal protease are well solved; the sample prepared by the method is complete to prepare, has a large quantity of protein isolating points, and is high in repeatability and clear in chromatogram; by virtue of detection of PDQuestsoftware, the number of the protein points is greater than 1,000; therefore, the method lays the foundation for further mass spectrometry.

The invention belongs to the field of molecular imaging, and discloses an in vivo tumor imaging target molecule and a specific probe thereof. The target molecule is sperm protein 17 (Sp17) which is expressed abnormally in a tumor cell; the specific probe comprises targeting agent and tracer; and the targeting agent is monoclonal antibody capable of being combined with Sp17 specific, and the tracer is fluorescent dye or radioactive nuclide. The probe can be combined with the specific of an in vivo tumor, has good imaging effect and good stability, and has good application prospect in preparing tumor molecular imaging diagnostic reagent.

The present invention provides compositions and methods useful for diagnosing, treating, and preventing cancer, particularly ovarian cancer and uterine cancer, based on the discovery that the oocyte specific protein, SAS1R (Sperm Acrosomal SLL-P1 Receptor), which is a sperm protein receptor, is also expressed in various cancers, including ovarian cancer and uterine cancer. Six SAS1R variants have been previously identified, and they are encompassed by the invention. The present invention further provides antibodies useful for targeting SAS1R expressing cells and for killing such cells.

The results herein identify Sp17 as a novel cancer-testis antigen in multiple myeloma. Sp17 recombinant protein was generated from E. coli. A CD8 predominant CTL line was generated that was able to lyse autologous targets in a Sp17-dependent HLA class I-restricted manner, using dendritic cells as the antigen-presenting cells and DOTAP to deliver the Sp17 protein to the dendritic cells. A combination of HLA-matched and mismatched antibody-enriched fresh myeloma tumor cells and myeloma cell lines were used as targets for the recombinant protein-propagated CTL. The findings of target cell lysis suggest that the Sp17 protein produced by Sp17+ tumor cells are processed and presented in vivo and that the CTL epitopes are presented in association with HLA class I molecules in a concentration and configuration recognized by recombinant protein-propagated CTL.

The invention discloses a sperm protein marker IZUMO2 closely related to the reproductive performance of a service boar and application of the sperm protein marker IZUMO2. The invention provides the novel sperm protein marker IZUMO2 which is closely related to the reproductive performance of the service boar, and the relative content of the sperm protein marker is significantly and positively correlated with the reproductive performance of the service boar; by detecting the relative content of a sperm protein IZUMO2, the sperm fertilizing capacity and the reproductive performance level of theservice boar can be judged, so that relatively accurate evaluation of the reproductive performance of the existing service boars can be realized; the protein marker can be utilized to accurately select and breed service boars with high reproductive performance in production, the service boars with poor reproductive performance can be removed and eliminated, and the improvements on the reproductiveproduction performance and economic benefits of a pig farm are facilitated. The sperm protein marker IZUMO2 is easy to use in production, the reproductive performance can be judged only through the detection of the content of the sperm protein marker IZUMO2, and no sampling damage and stress of the service boars is caused.

The invention provides an immunological contraception vaccine, including a DNA sequence coding a mouse sperm-specific membrane protein Izumo. The DNA sequence coding the mouse sperm-specific membrane protein Izumo is inserted into an eukaryotic expression vector which may be pCAGGS. The invention further provides a preparation method for the immunological contraception vaccine. After the nucleic acid vaccine is used to immunize a mouse, generation of a high-titer specific antibody against mouse Izumo in the mouse is induced, and reproductive capacity of the mouse is reduced. Moreover, the specific antibody generated by the mouse immunized by the nucleic acid vaccine can undergo a specific reaction with mouse sperm proteins and effectively inhibit egg-sperm binding of the mouse.

The invention provides a quantitative detection method of sperm surface Izumo1 protein. The method comprises the following steps: performing separation and pretreatment of a sperm, performing preliminary treatment of the sperm to expose the Izumo1 protein epitope of the sperm, marking the Izumo1 protein of the sperm by using fluorescein, and performing detection by using a flow cytometer. According to Tthe method provided by the invention, utilizes the flow cytometry is utilized to perform quantitative detection on the Izumo1 protein in the human sperm for the first time, directly realizes precise quantification of the Izumo1 protein is directly realized, is easy to realize standardization in operation is easy to realize, is quick and accurate effects are realized, and is high inthe precision is high. Compared with traditional methods for qualitative or semi-quantitative detection of Izumo1 protein, such as cell immunofluorescence and fluorescence in situ hybridization, the method provided by the invention has the advantages of convenient detection, high precision, high speed and the like; the method can be applied to quantitative detection of expression of sperm surface Izumo1 protein and is easy to popularize; and the detection result can be used for performing auxiliary evaluation on the fertilization ability of the sperm, so that the method is suitable for practical application.

The invention discloses a sperm piRNA and a sperm protein MitoPLD as biomarkers for detecting and predicting male infertility. The biomarkers include 10 piRNAs (mainly piR-1207 and piR-2107) and the MitoPLD protein. The piRNA combination and the MitoPLD protein can be used for detection and prediction of sperm motility. According to the sperm piRNA and the sperm protein MitoPLD as the biomarkers for detecting and predicting the male infertility, spermatozoa are more easily obtained without other tissues, and the sperm piRNA and the sperm protein MitoPLD as the biomarkers for detecting and predicting the male infertility belong to non-invasive examination; the sperm piRNA and the MitoPLD protein can reflect pathological and physiological conditions throughout spermatogenesis at a molecularlevel, and are used as an auxiliary detection index to improve accuracy of detection, and provide potential targets for treatment of male reproductive dysfunction.

The invention discloses a method for extracting protein from frozen cow semen. The method comprises the following steps of: unfreezing frozen semen and transferring to a centrifugal tube, centrifuging the unfrozen semen for 5 minutes at 4 DEG C and 1000*g, discarding the liquid supernatant; (2), adding 1ml of PBS (Phosphate Buffer Solution) which is pre-cooled to 4 DEG C, centrifuging for 5 minutes at 4 DEG C and 1000*g, repeating the process for three times; (3), adding 200 microliters of protein lysis solution to the precipitate and sufficiently dissolving the sample by volution; (4), stewing the solution for 5 minutes and centrifuging the solution for 30 minutes at 4 DEG C and 12000*g, wherein the obtained liquid supernatant is protein solution. The method for extracting protein from frozen cow semen disclosed by the invention is simple to operate, capable of laying a foundation for researching the semen freezing and protecting mechanism by molecular means in the future and capable of further improving the activity of the frozen semen.

The invention discloses an application of Matriptase. The Matriptase comprises a short intracellular segment, a transmembrane region, an SEA (sea urchin sperm protein, enteropeptidase, agrin domain) region, two CUB (Cls / Clr, urchin embryonic growth factor and bone morphogenetic protein-1) regions, four low density lipoprotein acceptor domains and a C-terminal trypsin-like protease region. The Matriptase is applied to diagnosis and treatment of chronic lymphocyte leukemia. By an experiment, bone marrows and peripheral blood specimens of normal people and leukemia sufferers are collected, and the detection result shows as follows: Matriptase mRNA and protein are highly expressed in the chronic lymphocyte leukemia sufferers, the expression level of Matriptase of the surface of lymphocyte can serve as a molecular marker for diagnosis of chronic lymphocyte leukemia, and the Matriptase can serve as a novel target for diagnosis and treatment of chronic lymphocyte leukemia.

The invention particularly relates to an application of RSPH9 as a diagnosis marker or a treatment target for oligoasthenozoospermia. Sperm proteins of normal and patients are extracted, bovine serumalbumin is used as a standard substance, and multiple groups of sperm proteins of severe application of RSPH9 as a diagnosis marker or a treatment target for oligoasthenozoospermia diseases are subjected to deep mass spectrometry by using high-resolution biomass spectrometry and a non-labeled quantitative proteomics method; the mass spectrum data are searched by using a protein quantitative analysis method, and the protein in the sperm is identified; and finally, normal and patient sperm proteomes are compared, and screening is conducted to obtain that the contents of RSPH9 in healthy and oligoasthenosperm samples are remarkably different. ROC analysis further verifies that RSPH9 has good specificity as a diagnostic marker, and is expected to be used as a diagnostic marker and a treatmenttarget for severe oligoasthenozoospermia.

The invention discloses a sperm protein marker SPACA4 closely related to the breeding boar reproductive performance and application thereof. According to the novel sperm protein marker SPACA4 closelyrelated to the breeding boar reproductive performance, the relative amount of the novel sperm protein marker SPACA4 and the breeding boar reproductive performance show significant positive correlation, by measuring the relative content of the sperm protein marker SPACA4 and using the relative content of the sperm protein marker SPACA4 for judging the sperm fertilizing capacity and the reproductiveperformance of breeding boars, the reproductive performance of the existing breeding boars can be accurately judged, in production, by means of the protein marker, breeding of the high-reproductive performance breeding boars can be conducted accurately, and the boars with poor reproductive performance are eliminated, so that the sperm protein marker SPACA4 is beneficial for improving the reproductive productivity and economic benefits of a pig farm; in production, use is convenient, only the content of the sperm protein marker SPACA4 needs to be detected, and breeding boar sampling damages orstress is avoided.

The invention discloses a sperm quality evaluation method, and belongs to the technical field of biological detection. According to the evaluation method, the proteasome activity in semen is taken as an index, the proteasome activity among different age groups, different sperm motility groups, different DNA fragment groups, different morphological rates and different head malformation rate groups is measured, the proteasome activity among different semen quality index groups is found to have no significant difference, but the higher the sperm proteasome activity expression is, the higher the sperm proteasome activity expression is, the higher the sperm proteasome activity expression is, the higher the sperm proteasome activity is. The fertilization rate and 2PN rate of conventional in-vitro fertilization are higher. Therefore, the sperm proteasome activity of a patient with unknown sterility or low conventional in-vitro fertilization rate can be considered and detected, and a basis is provided for clinically selecting conventional in-vitro fertilization or ICSI treatment.

The present invention provides compositions and methods useful for diagnosing, treating, and preventing cancer, particularly ovarian cancer and uterine cancer, based on the discovery that the oocyte specific protein, SAS1R (Sperm Acrosomal SLLP1 Receptor), which is a sperm protein receptor, is also expressed in various cancers, including ovarian cancer and uterine cancer. Six SAS1R variants have been previously identified, and they are encompassed by the invention. The present invention further provides antibodies useful for targeting SAS1R expressing cells and for killing such cells.

The present invention includes compositions and methods for treating a breast cancer cell, a metastatic breast cancer cell, or a triple negative breast cancer cell comprising: identifying a subject in need for treatment for a breast cancer cell, a metastatic breast cancer cell, or a triple negative breast cancer cell; and administering a therapeutically effective amount of a formulation that leads to the presentation of an immunogenic SP17 protein or peptide antigen on an antigen presenting cell to activate T cells that are SP17-specific T cells, wherein the SP17-specific T cells impair the growth of the breast cancer cell, the metastatic breast cancer cell, or the triple negative breast cancer cell growth.

The invention relates to immunocontraceptive synthetic peptide and immunocontraceptive chimeric peptide, wherein the amino acid sequence of the immunocontraceptive chimeric peptide is shown as SEQ ID:1 in a sequence table, the amino acid sequence of the immunocontraceptive synthetic peptide hESP103-107 is shown as SEQ ID:2 in the sequence table, and the amino acid sequence of the immunocontraceptive synthetic peptide hIzumo216-220 is shown as SEQ ID:3 in the sequence table. The invention also provides the application of the chimeric peptide, the synthetic peptide hESP103-107 or the synthetic peptide hIzumo216-220 as immunocontraceptive vaccines observed in contraception. The invention researches the chimeric peptide comprising sperm protein hESP, hIzumo and tNASP as well as the cross reacting antigens of UU (Ureaplasma Urealyticum) for the first time, and immunized mice can obtains stronger and reversible antifertility effect.

The invention provides a quantitative detection method of sperm surface Izumo1 protein. The method comprises the following steps: performing separation and pretreatment of a sperm, performing preliminary treatment of the sperm to expose the Izumo1 protein epitope of the sperm, marking the Izumo1 protein of the sperm by using fluorescein, and performing detection by using a flow cytometer. According to Tthe method provided by the invention, utilizes the flow cytometry is utilized to perform quantitative detection on the Izumo1 protein in the human sperm for the first time, directly realizes precise quantification of the Izumo1 protein is directly realized, is easy to realize standardization in operation is easy to realize, is quick and accurate effects are realized, and is high inthe precision is high. Compared with traditional methods for qualitative or semi-quantitative detection of Izumo1 protein, such as cell immunofluorescence and fluorescence in situ hybridization, the method provided by the invention has the advantages of convenient detection, high precision, high speed and the like; the method can be applied to quantitative detection of expression of sperm surface Izumo1 protein and is easy to popularize; and the detection result can be used for performing auxiliary evaluation on the fertilization ability of the sperm, so that the method is suitable for practical application.

The invention discloses a sperm protein marker SPACA4 closely related to the reproductive performance of boars and its application. The invention provides a new sperm protein marker SPACA4 which is closely related to the reproductive performance of boars, and its relative content is significantly positively correlated with the reproductive performance of boars. The fertilization ability and reproductive performance of pig semen can be used to accurately evaluate the reproductive performance of existing boars. In production, the protein marker can be used to more accurately select and breed high-reproductive boars. Elimination of boars with poor reproductive performance is conducive to improving the reproductive performance and economic benefits of pig farms. It is convenient to use in production, and it only needs to detect the content of the semen protein marker SPACA4, and will not cause boar sampling damage and stress.

The invention discloses a sperm protein marker IZUMO2 closely related to the reproductive performance of a service boar and application of the sperm protein marker IZUMO2. The invention provides the novel sperm protein marker IZUMO2 which is closely related to the reproductive performance of the service boar, and the relative content of the sperm protein marker is significantly and positively correlated with the reproductive performance of the service boar; by detecting the relative content of a sperm protein IZUMO2, the sperm fertilizing capacity and the reproductive performance level of theservice boar can be judged, so that relatively accurate evaluation of the reproductive performance of the existing service boars can be realized; the protein marker can be utilized to accurately select and breed service boars with high reproductive performance in production, the service boars with poor reproductive performance can be removed and eliminated, and the improvements on the reproductiveproduction performance and economic benefits of a pig farm are facilitated. The sperm protein marker IZUMO2 is easy to use in production, the reproductive performance can be judged only through the detection of the content of the sperm protein marker IZUMO2, and no sampling damage and stress of the service boars is caused.

The invention discloses a recombinant adeno-associated virus (rAAV) vector carrying a human sperm protein 17 (SP17) antigen gene and proves the application value of the recombinant adeno-associated virus vector. The rAAV vector is obtained by inserting an SP17 antigen gene into an AAV starting vector constructed by the inventor. The research proves that the virus vector of the rAAV can effectivelyconvey the carried SP17 antigen gene into a human mononuclear cell-dendritic cell line to stimulate effector cells of cellular immunity, that is, generation of SP17 antigen specific cytotoxic T lymphocytes (CTL). The experiment proves that the CTL induced by DC infected by the recombinant adeno-associated virus vector carrying the SP17 antigen gene can effectively split SP17 antigen positive malignant tumor cells. Therefore, the recombinant adeno-associated virus vector carrying the SP17 antigen gene or the related product thereof have practical values and can be used for preparing the SP17 antigen specific CTL; the SP17 antigen positive malignant tumor cells can be killed; and the targeted T cell immunotherapy on anti-SP17 antigen positive malignant tumors can be realized.

Cancer vaccines were demonstrated to be promising strategies for cancer treatment; however these strategies are limited by the paucity of target antigens that provoke an effective immune response. The Inventors of the present patent application propose that sperm fibrous sheet proteins constitute a new class of potential antigens for use in the development of effective, durable cancer vaccines. This hypothesis is supported by the expression of the sperm fibrous sheath protein, known as Sperm protein 17 (Sp17), which is detected in tumors of unrelated histological origin. It has the ability to induce T-cell based immune responses. The expression of the Sperm protein 17 (Sp17) in tumors of unrelated histological origin, and its' natural localization in the human sperm fibrous sheath (FS), led the Inventors to hypothesize that FS proteins might represent a new potential class of target antigens useful for developing cancer vaccines, as well as diagnostic and therapeutic treatments.

Methods of identifying cancer / testes antigens (CTAs) useful as cancer treatment targets are disclosed and claimed herein. The methods include identifying human sperm proteins to which patients diagnosed with solid or hematological malignancies have established a humoral immune response.

The invention provides a marker protein related to asthenospermia and an application of the marker protein, and particularly discovers that sulfur-sulfhydrylation modification of H3 protein in sperm protein participates to regulation and control of establishment and maintenance of sperm motility, and abnormal sulfur-sulfhydrylation modification of H3 participates in a pathological process of asthenospermia. The sperm motility can be obviously improved by improving the H3 sulfhydrylation level, so that the asthenozoospermia is prevented or treated.