180 results about "E2 protein" patented technology

Two Hepatitis C Virus envelope proteins (E1 and E2) are expressed without sialylation. Recombinant expression of these proteins in lower eukaryotes, or in mammalian cells in which terminal glycosylation is blocked, results in recombinant proteins which are more similar to native HCV glycoproteins. When isolated by GNA lectin affinity, the E1 and E2 proteins aggregate into virus-like particles.

The present invention provides modified hepatitis C virus genomic RNA, comprising nucleotide sequences of genomic RNA portions of two or more types of hepatitis C viruses, which comprises a 5′ untranslated region, a core protein coding sequence, an E1 protein coding sequence, a p7 protein coding sequence, an E2 protein coding sequence, an NS2 protein coding sequence, an NS3 protein coding sequence, an NS4A protein coding sequence, an NS4B protein coding sequence, an NS5A protein coding sequence, an NS5B protein coding sequence, and a 3′ untranslated region, and which can be autonomously replicated. In particular, the present invention relates to modified hepatitis C virus genomic RNA, which can be autonomously replicated by substitution of the RNA sequence portion encoding NS3, NS4, NS5A, and NS5B proteins of hepatitis C virus genomic RNA with a partial RNA sequence encoding NS3, NS4, NS5A, and NS5B proteins of a JFH1 strain shown in SEQ ID NO: 1.

The present invention provides systems for identifying anti-viral agents. In particular, the invention encompasses reagents and strategies for identifying agents that inhibit or disrupt key protein-protein interactions that are important in the life cycle of papillomaviruses. The invention allows identification, production, and/or use of agents that reduce or inhibit the replication of HPV by inhibiting (e.g., precluding, reversing, or disrupting) the formation of the E1-E2 protein-protein complex. The invention also provides specific inhibitory agents, pharmaceutical compositions, and methods of using these inhibitors and pharmaceutical compositions for inhibiting viral replication in vitro. Methods are also described for the treatment and prevention of HPV infections and HPV-related diseases in patients.

The invention provides a CSFV antibody detection system and a preparation method thereof. A coating antigen of the detection system contains CSFV E2 protein and Erns protein. The CSFV E2 protein and Erns protein are recombinant proteins expressed by eucaryon, correct spatial conformation and posttranslational modification process can be guaranteed, antigen is capable of effectively combining with the antibody in serum, and specificity, sensitivity and repeatability of detection can be increased. The system and the method can be sued for diagnosis of CSFV antibody in prevention and control of CSFV as well as immunization evaluation of a CSFV vaccine.

The invention discloses a recombinant virus for expressing swine fever virus E2 gene, and a preparation method and application thereof. According to the preference of the codon, synthetic CSFV 2.1-type immunogenic gene E2 and 1.1-type E2 gene are respectively arranged in baculovirus pH and p10 promoters to be expressed so as to obtain a recombinant baculovirus (CCTCC NO:V201338). The exogenous gene expression level is enhanced. After being used for immunizing Japanese white rabbits and pigs, the subunit vaccine prepared by the E2 protein expressed by the recombinant baculovirus can induce the organism to generate specific immune reaction and protect the organism from the attack of the swine fever virus; and differential diagnoses can be utilized to distinguish the vaccine immunized animal from the wild virus-infected animal, thereby being beneficial to eradication and purification of the eradicate swine fever virus in the pig farm.

The invention relates to a test paper card for detecting porcine transmissible gastroenteritis virus antibody, preparation and detection method thereof, and belongs to the field of immunological detection. The test paper card includes a jammed case and a test strip, and the test strip includes a bottom plate and sequentially lapped and pasted on the Absorbent pad, detection pad, binding pad and sample pad on the base plate; the detection pad is a nitrocellulose membrane with a quality control line C and a detection line T, the quality control line C is coated with goat anti-mouse monoclonal antibody, and the detection line T is Coated with inactivated porcine transmissible gastroenteritis virus; the binding pad is a glass cellulose membrane embedded with time-resolved fluorescent microsphere-labeled anti-E2 protein monoclonal antibody; the sample pad is dried glass after soaking in the sample treatment solution Cellulose film. The test paper card prepared by the invention has better stability and higher sensitivity, and can achieve the purpose of semi-quantitative detection through fluorescence signal analysis.

The invention discloses a monoclonal antibody against virulent strain E2 protein of classical swine fever virus and a hybridoma cell strain secreting the monoclonal antibody. The hybridoma cell strain is obtained by using hog cholera lapinized virus vaccine strain E2 protein expressed by Baculovirus as tolerogen, selecting Shimen strain E2 protein as immunogen, immunizing mouse by cyclophosphamide immunosuppression method, carrying out cell fusion, and sieving hybridoma cell strain capable of stably secreting monoclonal antibody against E2 protein. The monoclonal antibody can react with Shimen strain and can produce specific reaction with virulent strain of classical swine fever viruses of 1.1, 2.1, 2.2 and 2.3 gene sub-groups. The monoclonal antibody has neutralization activity and does not react with hog cholera lapinized virus vaccine strain, so that the monoclonal antibody can be used for differentiating virulent strain of classical swine fever virus and hog cholera lapinized virus vaccine strain, which establishes the foundation for establishing a method for differentiating wild virus infection of classical swine fever and vaccine immunity and for researching the molecular difference between CSFV virulent strain and mild strain.

The invention discloses preparation methods for CHO cell expressed recombinant bovine viral diarrhea virus protein E2 and a subunit vaccine and applications and belongs to the technical fields of animal vaccines and veterinary biologicals. The object of the invention is to provide a preparation method capable of industrially producing the bovine viral diarrhea virus recombinant subunit vaccine ona large scale. The preparation method for the recombinant subunit vaccine, provided by the invention, comprises the following steps: 1) cloning an eukaryotic expression vector containing a protein E2coding gene; 2) transfecting CHO cells, and performing selection, screening and acclimatizing to obtain suspending CHO cell strains, which stably and efficiently express the protein E2; 3) subjectingthe cell strains obtained in the step 2) to fermentation culture, and carrying out purification, so as to obtain recombinant protein E2; and 4) uniformly mixing the recombinant protein E2 and ISA 201VG thoroughly, thereby obtaining the recombinant subunit vaccine. According to the method provided by the invention, target protein can be obtained from cell culture supernatant, the yield reaches upto 500mg/L, the protein purification time is shortened, the vaccine production steps are simplified, and the vaccine production cost is greatly reduced.

The invention provides a classical swine fever E2 subunit vaccine and a preparation method thereof. The preparation method specifically comprises the steps that a Baculovirus Carrier Expression System is used for expressing a large amount of recombination classical swing fever E2 protein in insect cells, and the classical swine fever E2 subunit vaccine with a good immune effect is developed. The Bac-to-Bac Baculovirus Carrier Expression System is used for expressing the classical swine fever E2 protein. The classical swine fever E2 protein and an adjuvant 201R are emulsified into the novel subunit vaccine, and by means of piglet immune challenge tests, it is verified that the subunit vaccine has a good immune protection effect.

The invention discloses preparation of a neutralizing monoclonal antibody of an anti-hepatitis C virus and application thereof. E2 protein is utilized to strengthen immunity after mice are immunized by pVAX-CpG-N2N8; spleen of the mice with the highest valence aiming at the E2 antibody is taken as an antigen-sensitized B cell and fused with a myeloma cell SP2/0 strain, and the fused cell is screened in HAT culture medium, so as to obtain the fused cell, and the fused grows and is cloned to obtain the hybridoma cell strain generating the monoclonal antibody disclosed by the invention. The neutralizing monoclonal antibody of the anti-hepatitis C virus E2 disclosed by the invention is generated by the hybridoma cell strain; epitope combined with the monoclonal antibody is at F550GCTWMNSTGFTKVCGAPPCVIG572 part of E2 glycoprotein. The neutralizing monoclonal antibody of the anti-hepatitis C virus disclosed by the invention has good capability of neutralizing HCV infection, can be used as a hepatitis C virus therapeutic antibody, and has a great application prospect in hepatitis C diagnosis and treatment.

The invention discloses a recombinant swine fever virus E2 protein swine source monoclonal antibody and a preparation method and application thereof. Firstly, the recombinant swine fever virus E2 protein swine source monoclonal antibody is disclosed, an amino acid sequence of a heavy chain is shown in SEQ ID NO.1, and an amino acid sequence of a light chain is shown in SEQ ID NO.2. The invention also discloses a suspension HEK293 cell line which can stably express the recombinant swine fever virus E2 protein swine source monoclonal antibody. The recombinant swine fever virus E2 protein swine source monoclonal antibody has the advantages that the coding genes of the heavy chain and light chain are cloned into an eukaryotic expression vector, and the stable and high-efficiency expression ofthe recombinant swine fever virus E2 protein swine source monoclonal antibody is realized by the suspension HEK293 cell line; the reactogenicity and neutralizing activity are good, and the important development value is realized in development of novel swine fever virus diagnosing and treating preparations.

The invention mainly relates to a polypeptide sequence combined with bovine viral diarrhea E2 protein and application of the polypeptide sequence. The polypeptide sequence is KRLREL and is a linear combined polypeptide, the polypeptide sequence can be prolonged and modified by taking the polypeptide sequence as a core, and modification materials can be but not limited to nano materials, fluorescent materials, enzymes, biotin and specific protein. By adopting a phage peptide library screening technique, the expressed and purified bovine viral diarrhea E2 protein can be screened for multiple rounds, and polypeptide clone strains which can be specifically combined with the bovine viral diarrhea E2 protein can be ultimately obtained; the polypeptide clone strains can be selected for sequencing, the core sequence of the polypeptide is KRLREL, the manually synthesized polypeptide used for ELISA combination experiment shows that the synthesized polypeptide can be well combined with the bovine viral diarrhea E2 protein; the process is simple, and the operation is relatively convenient compared with the operation that expression protein is manually expressed and is further immunized to obtain a protein antibody; by marking the polypeptide, a kit or a test paper bar (card) for quantitative and qualitative detection on the bovine viral diarrhea E2 protein can be rapidly produced.

The invention discloses a test paper for rapidly detecting swine fever antibody and a preparation method thereof. The inventive test paper consists of a sample pad (4), a glass fiber member (3), a cellulose nitrate membrane (2), a water absorbent pad (1) and a support (5); wherein the cellulose nitrate membrane contains a detection line (6) coated by swine fever virus E2 protein and a reference line (7) coated with rabbit anti swine fever antibody; and the glass fiber member is combined with swine fever virus E2 protein marked by colloidal gold. The test paper can rapidly detect possibly present swine fever antibody in a sample to achieve rapid detection and timely epidemic control, thus creating favorable conditions for further separation and identification. The test paper has the advantages of convenient, rapid and easy usage, clear result, easy generalization, and no need for special instrument and equipment as well as professionals, and is suitable for large-batch onsite detection for base layer and for sudden events. The test paper is suitable for epidemic inquisition and performs assistant effect on swine fever virus infection diagnosis.

The invention relates to a kit for quickly detecting a swine fever antibody. In the method, a pair of specific primers is designed, a relatively conservative gene sequence is cloned, an antigen aiming at a swine fever E2 protein is expressed through a pronucleus expression technology, and, on the basis, the kit containing an enzyme linked plate coated with a high-purity and high activity swine fever virus specificity antigen, an enzyme conjugate of rabbit-anti-swine monoclonal antibody containing an HRP (Horseradish Peroxidase) marker, a TMB (Tetramethylbenzidine) color developing liquid and the like is prepared. The kit can quickly detect the swine fever antibody in blood serum or blood plasma and has strong specificity and high sensitivity.

The invention discloses a swine fever-porcine circovirus combined subunit vaccine, as well as a preparation method and an application thereof, and belongs to the technical fields of animal vaccines and veterinary biological products. The vaccine is prepared from swine fever virus E2 protein, porcine circovirus type 2 cap protein and a pharmaceutically acceptable adjuvant. The preparation method comprises the following steps: (1) preparing swine fever virus E2 protein and porcine circovirus type 2 cap protein; (2) preparing an antigen solution from the swine fever virus E2 protein and porcine circovirus type 2 cap protein prepared in the step (1); and (3) mixing and stirring the antigen solution and Montanide GEL 01PR adjuvant in a mass ratio of 10:1. The vaccine has the advantages of strong immunogenicity, good safety, no immune interference and the like, can be used for preventing potential biological safety hazard for virus variation and fundamentally purifying swine fever virus and porcine circovirus type 2, and can achieve the effect of dual prevention with one injection by immunizing the vaccine to achieve the aims of saving time, labor and cost.

A glycoprotein E2 of classical swine fever virus (CSFV) expressed in a recombinant yeast system. The recombinant E2 protein (yE2) is able to form a homodimer, exhibits glycosylation conformation and possesses correct immunogenicity. An anti-CSFV vaccine can be provided with yE2 as a major active ingredient to induce high titers of neutralizing antibody in vaccinated pigs, and to induce a protection against CSFV infection.

The present invention provides CD4+ T-cell epitopes in E6, E7 and E2 proteins from various strains of human papillomavirus (HPV). In some preferred embodiments, the present invention provides means for the development of HPV vaccines, in particular multivalent vaccines for the prevention of infection with high-risk HPV strains. In additional embodiments, the present invention provides means for the development of therapeutic vaccines against high-risk HPV types that prevent the development of benign and/or malignant tumors in infected individuals. The present invention further provides epitopes suitable for use in prophylactic and therapeutic vaccines.

The use of a compound of formula (II):

or its enantiomers or diastereoisomers thereof, or salts or pharmaceutically-acceptable esters thereof, in the treatment or prevention of a papilloma virus infection, particularly human papilloma virus in a mammal, wherein R¹¹; X₄; X₅; X₆; R¹³; R¹⁴; W; Z; Y; T; and R¹⁸ are defined herein.

The present invention also provides novel compounds, pharmaceutical compositions and methods for using these compounds and compositions in the treatment or prevention of papilloma virus infection. More particularly, the present invention provides compounds, compositions and methods for inhibiting papilloma virus DNA replication by interfering with the E1-E2 protein-protein interaction essential for viral DNA replication.