13 results about "Pseudovirion" patented technology

The invention discloses an anti-Crimean-Congo hemorrhagic fever virus (CCHFV) neutralizing antibody N013 and application thereof in diagnosis, treatment and / or prevention of Crimean-Congo hemorrhagic fever, and the antibody has a unique CDR region and can be combined with the Gc protein of the CCHFV with high affinity. In a virus neutralization experiment, the antibody N013 disclosed by the invention has strong neutralization activity on CCHFV pseudovirus particles, the IC50 value is 26.63 ng / mL, and the antibody N013 has a wide application prospect in treatment of Crimean-Congo hemorrhagic fever.

The invention belongs to the technical field of pseudovirus detection, particularly relates to a method for evaluating a neutralizing antibody by using pseudovirus particles, and aims to improve the stability and sensitivity of a pseudovirus system in neutralizing antibody screening and cell membrane fusion analysis. The pseudovirus construction system comprises an envelope protein expression plasmid, a fluorescent reporter plasmid containing a marker sequence and an auxiliary packaging plasmid, the structural design of the envelope protein is optimized by adopting a positioning signal domain and a non-repetitive flexible peptide fragment, and a standard transfection and infection process is adopted, so that the pseudovirus is obtained. The accurate evaluation on the neutralizing antibody reaction under the dynamic culture condition is realized. Experimental results show that the construction system is superior to a traditional structure in the aspects of infection efficiency, fluorescence expression intensity, fusion integrity, neutralization inhibition effect and the like, and is suitable for optimization design of a pseudovirus platform and function verification of a neutralizing antibody.

The present invention discloses an anti-Crimean-Congo hemorrhagic fever virus (CCHFV) neutralizing antibody N015 and its use in the diagnosis, treatment and / or prevention of Crimean-Congo hemorrhagic fever, the antibody having a unique CDR region capable of binding to the Gc protein of CCHFV with high affinity. In a virus neutralization experiment, the antibody N015 disclosed by the invention has strong neutralization activity on CCHFV pseudovirus particles, the IC50 value is 53.39 ng / mL, and the antibody N015 has a wide application prospect in treatment of Crimean-Congo hemorrhagic fever.

According to the invention, a Chinese forest encephalitis virus inactivated vaccine is adopted to produce a strain'senzheng 'strain gene sequence, a structural protein preM-E gene sequence of the strain is amplified, an HIV-1 lentivirus packaging system is utilized, preM-E is used as a pseudovirus particle envelope protein, green fluorescent protein (EGFP) or luciferase reporter gene (Luciferae) is used as a detection indication signal to prepare the pseudovirus, and a BHK21 cell is used as a target cell; the pseudovirus-BHK21 cell system is used for replacing live virus operation experiments such as vaccine potency evaluation, neutralizing antibody detection (animal method or plaque reduction neutralization test) and the like, the biosafety operation environment is degraded from BSL-3 to BSL-2 by using pseudoviruses, the detection period is shortened, and animal experiments are reduced.

A method for preparing an immunogenic composition, which comprises treating HCV pseudoparticles with α2-3,6,8,9 neuraminidase A to generate the immunogenic composition. In addition, the immunogenic compositions are made in vaccines for eliciting an immune response to HCV in a subject.

The present invention relates to a type I or II transmembrane fusion protein comprising, successively:a) optionally, a signal peptide;b) a protein or a peptide of interest;c) a coiled-coil domain; andd) a domain for anchoring in the plasma membrane, consisting of a transmembrane segment and a cytosolic segment.It also relates to the virus-like particles obtained with this fusion protein.

The invention provides a reverse pseudovirus particle based on a receptor as well as a preparation method and application of the reverse pseudovirus particle, and belongs to the technical field of biological pharmacy. A transmembrane protein receptor derived from a host is integrated on an envelope of the pseudovirus particle, and an extracellular structural domain of the transmembrane protein receptor faces the outside of the virus particle, so that the pseudovirus particle can specifically recognize and combine target cells expressing corresponding virus envelope protein, and the pseudovirus particle is mediated to enter the target cells. The skeleton of the pseudovirus particle is selected from one of VSV [delta] G, HIV or MLV. The transmembrane protein receptor is I-type transmembrane protein, and is preferably selected from ACE2, DPP4 or APN. According to the invention, a prefabricated and extensible reverse receptor pseudovirus detection system is constructed, multi-receptor cross-species function evaluation and rapid new coronavirus receptor identification are realized, and an efficient and universal platform is provided for cross-species propagation monitoring, drug screening and vaccine evaluation.

The invention discloses an application of a marketed drug AZD-9291 (Osimertinib, osimertinib) in resisting infection of alphaviruses such as ONNV (O'nyong-nyong virus) and the like, and relates to the technical field of drugs, in particular to an application of the drug AZD-9291 (Osimertinib, osimertinib) in resisting infection of alphaviruses such as ONNV (O'nyong-nyong virus) and the like. Experimental results show that the AZD-9291 can be used for effectively inhibiting the infection of four pseudovirus particles, namely ONNV (ONNV), CHIKV (Chikungunya virus), EEEV (Eastern equine encephalitis virus) and SFV (Semliki Forest Virus), on host cells, and the AZD-9291 can be used for effectively inhibiting the infection of the four pseudovirus particles, namely ONNV, CHIKV, Chikungunya virus, EEEV, Eastern equine encephalitis virus, and SFV (Semliki Forest Virus, forest encephalitis virus). The CC50 of AZD-9291 in HEK293T cells is determined to be 53.3 [mu] M, then the inhibition effect of the compound on ONNV pseudoviruses is determined, the IC50 of the compound is 1.41 [mu] M, and meanwhile, the compound is found to have no inhibition effect on infection of rVSV [delta] G-G control pseudoviruses. Subsequently, the inhibition effect of AZD-9291 on other alphaviruses such as CHIKV, EEEV and SFV in HEK293T cells is determined, and the IC50 (half maximal inhibitory concentration) of AZD-9291 is 3.74 mu M, 5.18 mu M and 3.47 mu M respectively. The research proves that AZD-9291 can be used as a broad-spectrum candidate drug for inhibiting infection of alphaviruses such as ONNV, CHIKV and the like, and new application and new prospect of AZD-9291 as a clinical alphavirus infection treatment drug and preparation are shown.

The invention discloses a veterinary fluorescent PCR (polymerase chain reaction) detection kit and a detection method, and relates to the technical field of biological detection.The kit comprises a reaction solution A, a reaction solution B and a reaction solution C, the reaction solution A comprises a 5xBuffer providing a reaction buffer system and dNTPs, a specific primer pair aiming at PEDV, RV, PDCoV and TGEV and a probe; the enzyme mixture B comprises a hot start Taq DNA polymerase, a reverse transcriptase, a UNG enzyme (used for preventing pollution) and a stabilizer; the positive control is pseudovirus particles or in-vitro transcription RNA containing target gene fragment sequences of PEDV, RV, PDCoV and TGEV and is dissolved in RNA preserving fluid, PCR amplification is monitored in real time through fluorescence signals, and virus nucleic acid is quantitatively detected. By designing specific primers and probes and targeting gene sequences of PDCoV, TGEV, PEDV and PoRV, the accuracy and sensitivity of detection are ensured, and the kit has great application value and popularization prospect in quarantine, epidemic disease prevention and control and purification.

This invention discloses a method for detecting viral or pseudoviral cell infection, comprising: applying a sinusoidal voltage to an attached membrane in patch-clamp mode and detecting the corresponding membrane current; inducing viral or pseudoviral cell infection by binding of viruses or pseudoviruses in the electrode fluid to receptors on the attached membrane; performing phase discrimination demodulation on the corresponding membrane current signal using the phase of the input sinusoidal voltage as a standard to obtain the imaginary and real parts of the membrane current and calibrating and standardizing them in real time; calculating the membrane capacitance of the attached membrane based on the real and imaginary parts of the current signal, based on the calibration and standardization; identifying viral or pseudoviral cell infection events and their modes based on the dynamic characteristics and polarity of the membrane capacitance changes; and estimating the size of the virus or pseudovirus based on the magnitude of the membrane capacitance changes. This invention enables real-time, high-precision detection of various viruses or pseudoviruses infecting host cells via envelope-cell membrane fusion or endocytosis in acutely isolated cells, cultured cells, and isolated tissue cells.

This invention provides an in vitro packaging method for ring virus pseudovirus particles, belonging to the field of molecular biology technology. The method involves eukaryotic expression of a ring virus capsid expression plasmid. Ring virus pseudovirus particles can serve as a safe and efficient novel viral vector for gene therapy and basic research. The technical method of this invention will promote basic research on ring viruses and related vector-based applications.

The present invention discloses an anti-Crimean-Congo hemorrhagic fever virus (CCHFV) neutralizing antibody N025 and its use in the diagnosis, treatment and / or prevention of Crimean-Congo hemorrhagic fever, the antibody having a unique CDR region capable of binding to the Gc protein of CCHFV with high affinity. In a virus neutralization experiment, the antibody N025 disclosed by the invention has strong neutralization activity on CCHFV pseudovirus particles, the IC50 value is 22.74 ng / mL, and the antibody N025 has a wide application prospect in treatment of Crimean-Congo hemorrhagic fever.