1141 results about "Capsid" patented technology

The disclosure relates to compositions and methods for the preparation, use, and / or formulation of adeno-associated virus capsid protein variants.

The invention discloses a high-pathogenicity porcine reproductive and respiratory syndrome virus mRNA vaccine composition and application thereof. The vaccine composition is selected from any one of the following combinations: (1) an mRNA vaccine composition consisting of a PRRSV GP3 mRNA (messenger ribonucleic acid), a PRRSV GP4 mRNA (messenger ribonucleic acid) and a PRRSV GP5 mRNA (messenger ribonucleic acid); or (2) an mRNA vaccine composition consisting of a PRRSV GP5 mRNA, a PRRSV membrane protein (M) mRNA and a PRRSV nucleocapsid protein (N) mRNA. According to the invention, a high-pathogenicity PRRSV JX-A1 strain is used as a template, and mRNAs capable of synthesizing a plurality of fusion proteins GP345, GP5M and GP5MN and single structural proteins GP3, GP4, GP5, M and N are respectively constructed. After liposome encapsulation, mice are inoculated for immunological evaluation, and results show that humoral immunity and cellular immunity levels of a single mRNA-LNPs mixed group are higher than those of recombinant mRNA-LNPs, and GP5 + M + N and GP3 + 4 + 5 have relatively best effects. The invention provides a new technical means for preventing PRRSV (Porcine Reproductive and Respiratory Syndrome Virus).

The invention discloses an oligopeptide composition for inhibiting inclusion body formation and promoting Gag-Like protein nanoparticle self-assembly, the oligopeptide composition comprises a C-terminal oligopeptide and an N-terminal oligopeptide, the amino acid sequence of the N-terminal oligopeptide is as shown in SEQ ID NO: 1, and the amino acid sequence of the C-terminal oligopeptide is as shown in SEQ ID NO: 2. The oligopeptide composition can effectively avoid formation of a prokaryotic expression protein inclusion body and / or promote dissolution of prokaryotic expression protein, the oligopeptide composition is further utilized to be combined with HIV-1p24 and human endogenous retrovirus K capsid protein CA, nanoparticles are prepared respectively, and the oligopeptide composition can be used as a carrier for antigen surface display or drug transportation. The nanoparticles formed by the HIV-1 capsid protein p24 and the human HERV K CA have cavities which are obviously larger than those of ferritin, so that the nanoparticles have higher loading capacity; as an antigen display platform, the antibody also has good stability and wide applicability.

The invention provides a nano antibody and application thereof, a variable region in an amino acid sequence of the nano antibody comprises a complementarity determining region CDR and a framework region FR, the complementarity determining region CDR comprises a complementarity determining region CDR1, a complementarity determining region CDR2 and a complementarity determining region CDR3, the CDR3 is RWPDXY family, and binding sites of the CDR3 and AAV capsid are W99, D101 and Y103. The nano antibody provided by the invention has very high affinity and activity, and can specifically recognize and combine various serotypes of AAV, and an adsorbent prepared from the nano antibody has very strong adsorption capacity on AAV, can be applied to AAV affinity chromatography, and is beneficial to industrial application of an AAV affinity chromatography column. In addition, the method can also be applied to the field of AAV detection, and can detect empty capsid and virions together.

The disclosure is directed in part to variant capsid polypeptides that can be used to deliver payloads.

Methods are disclosed for delivering a composition comprising an adeno-associated virus (AAV) to one or more kidney cell types in a subject with or without immunity against AAV, including locally delivering to the kidney of the subject an adeno-associated virus (AAV) comprising a genome and an AAV capsid protein via renal pelvic injection, wherein the renal pelvic injection comprises direct renal pelvic injection or retrograde transureteral renal pelvic injection. Methods are also disclosed for treating a kidney disorder in a subject with or without immunity against AAV, comprising locally delivering to the kidney of the subject via renal pelvic injection, including direct renal pelvic injection and retrograde transureteral renal pelvic injection, an adeno-associated virus (AAV) comprising a genome and an AAV capsid protein, wherein the genome encodes a therapeutic agent that treats the kidney disorder in the subject. The AAV can be AAVKP1, AAVKP2, AAVKP3, AAVDJ, AAV2G9, AAV2.7m8, AAV3, AAVLK03, or AAVShH10.

The disclosure is directed in part to variant capsid polypeptides that can be used to deliver payloads.

Provided herein are compositions including brain, brain microvascular endothelial cell-targeting peptides and / or blood-brain barrier crossing (BBB) tissue-targeting peptides linked thereto or inserted in a targeting protein of a recombinant vector having at least one exogenous peptide comprising Xn – n-mer – Xm. Compositions providing such conjugates, targeting peptides, or recombinant vectors having an engineered capsid or envelope protein are provided as are uses thereof.

The invention belongs to the technical field of biological medicine, and particularly relates to a preparation method of viroid particles displaying scFv antibodies, a product of the viroid particles and application of the viroid particles. The preparation method provided by the invention comprises the following steps: constructing a specific targeting recombinant plasmid, wherein the specific targeting recombinant plasmid is obtained by connecting a vector for expressing virus capsid protein and a coding gene for expressing an scFv antibody display structure through homologous recombination. The vector for expressing the virus capsid protein comprises a coding gene of VSV-G signal peptide, a transmembrane region and an intracellular region, and is as shown in SEQ ID NO. 2. And the coding gene for expressing the scFv antibody display structure is shown as SEQ ID NO. 3-5. According to the preparation method disclosed by the invention, the virus-like particles of which the capsid is a modular component of a specific targeting immune cell are constructed based on a lentivirus system, and 50-90% of infection efficiency can be realized in the immune cell.

The invention discloses a lytic bacteriophage for preventing and controlling vibrio in an aquaculture environment. The bacteriophage vBValMR42H belongs to a muscle tail bacteriophage and is insensitive to chloroform, the capsid of the bacteriophage vBValMR42H is free of lipid substances, and the preservation number is GDMCC No: 67434-B1. The bacteriophage has the characteristics of high adsorption speed, short incubation period and high cracking amount, keeps high activity in a wide range of temperature (4-55 DEG C) and pH (2-11), and has strong environmental adaptability. Genome analysis shows that the bacteriophage does not contain virulence factors, antibiotic resistance genes and lyogen related genes, and is high in biological safety. Physical development analysis shows that the method can be divided into a new genus. The bacteriophage R42H has the advantages of high splitting efficiency, good environmental stability, strong specificity, gene safety and the like, and can be used as an ideal biological prevention and control agent for preventing and controlling vibrio alginolyticus diseases in aquaculture.

The present invention belongs to the field of genetic engineering and biotechnology, and relates to the screening, construction and application of a modified type of AAV-8 serotype for gene targeting and expression. The modified type of the AAV-8 serotype of the present invention inserts 10 amino acid sequences between the 590th amino acid and the 591st amino acid of the AAV-8 serotype capsid protein sequence as shown in SEQ ID NO.2, wherein the 3rd to 9th amino acids are amino acid sequences obtained by screening. The modified serotype of the present invention targets each layer of the retina, and the fluorescence effect after infection is stronger, and it can be clearly and intuitively observed that the exogenous gene can be efficiently expressed in each layer of the retina of the targeted tissue, including the retinal ganglion cell layer, the inner nuclear layer, the outer nuclear layer, the photoreceptor cell layer and the retinal pigment epithelial cell layer, and has better and more targeted tissue expression than the wild-type serotype when the same amount of virus is used.

The present invention belongs to the technical field of biological prevention and treatment for viral diseases. Disclosed in the present invention are a tomato disease-resistant gene mutant, and the use thereof in the prevention and treatment of ToBRFV. It is found in the present invention that mutating the nucleotide at position 1927 of the coding region sequence of tomato Tm-22 gene from G to A, or mutating the amino acid at position 643 of the LRR domain of a protein that is encoded by tomato Tm-22 gene from glycine to arginine can remarkably reduce the accumulation level of ToBRFV capsid protein (CP). The gene (named Tm-22-Mut5) can serve as a novel ToBRFV resistant gene, and also retains the resistance to TMV, ToMV and ToMMV. The Tm-22-Mut5 and a pre-screened Tm-22-Mut3-1 mutant (tyrosine at position 767 of the LRR domain thereof is mutated to phenylalanine) undergo combinatorial mutagenesis to obtain a mutant Tm-22-Mut6. Analysis shows that the Tm-22-Mut6 can remarkably reduce the accumulation level of the ToBRFV capsid protein. Compared with the pFGCTm-22-Mut3-1 mutant obtained by screening in the previous research and the newly obtained mutant Tm-22-Mut5, the Tm-22-Mut6 has further improved resistance to ToBRFV.

The invention discloses a method for preparing human metapneumovirus nucleocapsid protein based on an insect-baculovirus system. Comprising the following steps: 1) constructing a recombinant vector with a human metapneumovirus nucleocapsid protein coding sequence: sequentially connecting a sequence for promoting secretory expression, the human metapneumovirus nucleocapsid protein coding sequence and a tag sequence from 5'to 3 'ends in the recombinant vector; 2) converting the recombinant vector with the human metapneumovirus nucleocapsid protein coding sequence constructed in the step 1) into a competent cell containing a baculovirus genome plasmid Bacmid to obtain a recombinant baculovirus genome plasmid; and 3) transfecting insect cells with the recombinant baculovirus genome plasmid obtained in the step 2), culturing and purifying to obtain the human metapneumovirus nucleocapsid protein. The invention provides the preparation method of the human metapneumovirus nucleocapsid protein, and a material support is provided for subsequent further research.

The invention belongs to the technical field of gene engineering and biology, discloses a modified adeno-associated virus AAV-8 serotype for retina wide targeting and expression, and further relates to construction and application of the modified type. 10 amino acid sequences as shown in SEQ ID NO.3 are inserted between the 590th amino acid and the 591th amino acid of an AAV-8 serotype capsid protein sequence as shown in SEQ ID NO.2, the first, second and tenth amino acids are protected amino acids, and the third to ninth amino acids are amino acid sequences obtained by screening. The modified serotype disclosed by the invention is stronger in fluorescence effect after infection and can be obviously and intuitively observed, an exogenous gene can be widely and efficiently expressed in a specific targeted tissue retina whole region, and the expression efficiency is greatly improved under the condition of using the same virus amount. And compared with a wild serotype, the serotype has better and wider targeted tissue expression and less non-targeted tissue expression.

The present letter invention provides an engineered AAV capsid in which at least one protein on the capsid is modified to include an n-mer motif that promotes transduction of the capsid into the central nervous system (CNS) by interaction with a transferrin receptor. Further embodiments provide a carrier system comprising one or more carriers encoding an AAV capsid and a method of delivering a cargo to a CNS. The methods comprise administering an AAV capsid according to embodiments described herein in vivo or in vitro, and the AVV capsid comprises one or more carrier molecules.

The disclosure relates to compositions and methods for the preparation, use, and / or formulation of adeno-associated virus capsid protein variants.

The invention relates to the field of aquaculture, in particular to a preparation method and application of a larimichthys crocea iridovirus recombinant live vector vaccine. According to the invention, replication-deficient type 5 adenovirus is selected as a target gene expression vector, a recombinant adenovirus vector comprising main capsid protein of iridovirus is constructed, recombinant virus is obtained, and the safe and effective large yellow croaker iridovirus disease recombinant live vector vaccine is created. According to the vaccine, the larimichthys crocea is immunized in a soaking or bait feeding mode, the CPM of the larimichthys crocea immunized by the vaccine is 12.0%, and the CPM of the fish without being immunized by the vaccine is 62.0%. In addition, the RPS of the inoculated fish and the RPS of the empty carrier control group fish are 80.65% and 9.68% respectively. The method is at a leading level in the field of pseudosciaena crocea biomedicine at present, and creativity is remarkable.

The disclosure relates to compositions and methods for the preparation, use, and / or formulation of adeno-associated virus capsid protein variants.

The present disclosure relates to dosing regimens of an HIV capsid inhibitor and methods for the treatment or prevention of a human immunodeficiency virus (HIV) infection in a patient.

The invention discloses a panda-derived feline panleucopenia virus RAA-CRISPRCas12a detection method, and belongs to the technical field of virus detection, and the panda-derived feline panleucopenia virus RAA-CRISPRCas12a detection method comprises the following steps: S1, primer and probe design; s2, screening an RAA amplification primer; s3, RAA reaction condition optimization; s4, performing RAA sensitivity and specificity verification; s5, preparing crRNA (Complementary Ribonucleic Acid); s6, optimal crRNA screening is carried out; s7, optimizing the RAA-CRISPR / Cas12a (Reverse Amplified Architecture- S8, the specificity and the sensitivity of the RAA-CRISPR / Cas12a are verified; s9, clinical sample detection; according to the invention, by elaborately designing a primer and a probe aiming at the capsid protein VP2 gene of the feline parvovirus (FPV), fully utilizing the advantages of recombinase-mediated amplification (RAA) and a CRISPR / Cas12a system, and comprehensively optimizing reaction conditions and components of the system, the detection method has remarkable advantages, can quickly complete the detection process under the condition of constant temperature of 37 DEG C, is extremely high in sensitivity, and can be used for rapidly detecting the feline parvovirus. The lowest detection limit can reach 1 * 10 < 1 > copies / mu L, the specificity is high, and the coincidence rate with a real-time fluorescent quantitative PCR method is as high as 100%.

The present invention relates to a viral gene therapy vector for removing hepatitis B viruses, a preparation method therefor, and the use thereof, aiming to solve the problem in which existing viral gene therapy vectors can only inhibit the replication of hepatitis B viruses, but cannot remove virus cccDNA. The vector comprises a promoter, an intron, an enhancer, a nuclear localization signal (NLS) functional domain encoding gene of PGLYRP2 protein, an HBV DNA binding domain encoding gene of the PGLYRP2 protein, a secretory signal peptide functional domain encoding gene of the PGLYRP2 protein, an HBV nucleocapsid-binding functional domain encoding gene of the PGLYRP2 protein, and an IRES fragment. The viral gene therapy vector can remarkably promote the removal of HBVs in the liver of mice and is used for treatment of hepatitis B virus infection.

The disclosure relates to compositions and methods for the preparation, use, and / or formulation of adeno-associated virus capsid protein variants.

The invention relates to the field of biological medicine, in particular to an adeno-associated virus as well as a preparation method and application thereof. The capsid protein of the adeno-associated virus AAV-ACM205 provided by the invention comprises a wild type AAV2 capsid protein or a variant of the wild type AAV2 capsid protein, the wild type AAV2 capsid protein or the variant of the wild type AAV2 capsid protein comprises peptide fragments with amino acid sequences as shown in SEQ ID NO.6 and SEQ ID NO.7, and a random peptide fragment is inserted between the SEQ ID NO.6 and the SEQ ID NO.7. The adeno-associated virus vector AAV-ACM205 provided by the invention can mediate a target gene to be specifically and efficiently expressed in hair cells in cochlea of an adult mouse under the condition of semicircular canal injection; the adeno-associated virus vector AAV-ACM205 mediated target gene expression is more flexible and efficient than that of a traditional transgenic method, is convenient to apply and low in cost, and has a wide prospect in ear disease gene therapy.

The invention relates to the technical field of biological genetic engineering, in particular to construction of recombinant lactobacillus plantarum of an African swine fever virus antigen, and the method comprises the following steps: constructing a lactobacillus plantarum shuttle vector containing different ASFV antigen genes; and referring to a genome sequence of a Chinese isolate CADCHN09 of ASFV disclosed in GenBank. The method has the advantages that an inducer is not needed, antigen protein is efficiently expressed, the dendritic cell (DC) binding peptide for enhancing immune cell uptake and processing is subjected to fusion expression, the oral immunization mode is simple, mucosal immunity is stimulated, and ASFV infection is effectively prevented and controlled; probiotics, namely lactobacillus plantarum, which naturally exist in animal intestinal tracts and can be colonized and copied in digestive tracts for a long time are used as a carrier; key epitopes of proteins such as nucleocapsid protein p72, outer envelope protein CD2v, inner envelope protein p54, p22, pE199L and pE248R, which play an important role in adsorption, internalization, endocytosis and membrane fusion of ASFV, are preferably fused with dendritic cell binding peptide and then are displayed on the surface of lactobacillus plantarum.

The present invention relates to an adeno-associated virus (AAV) capsid protein variant having an amino acid mutation at the 328th to 331st amino acid residues of the AAV capsid protein relative to the 328th to 331st amino acid residues of the parent AAV capsid protein, for example, an amino acid sequence as shown in SEQ ID NO: 4 to replace the 328th to 331st amino acid residues of the parent AAV capsid protein wherein the amino acid position is numbered according to SEQ ID NO: 5. The invention also relates to an isolated nucleic acid encoding the AAV capsid protein variant, a nucleic acid vector comprising the same, and a host cell. The present invention also relates to a recombinant adeno-associated virus (rAAV) vector comprising said capsid protein variant, a pharmaceutical composition comprising said rAAV vector, and uses thereof in the treatment or prevention of skeletal muscle diseases.

The invention discloses a type 2 GAstV capsid protein monoclonal antibody and an antigen epitope peptide of the type 2 GAstV capsid protein monoclonal antibody. The monoclonal antibody is prepared by immunizing a mouse with capsid protein GAstV Cap of the type 2 goose astrovirus as an antigen, and is named as 1F1; the monoclonal antibody 1F1 comprises a heavy chain variable region and a light chain variable region, and the amino acid sequences of the heavy chain variable region and the light chain variable region are sequentially shown as SEQ ID No.3 and SEQ ID No.5. In order to determine the B cell antigen epitope of the screened monoclonal antibody, the invention also constructs a GAstV Cap truncated fusion protein, and the antigen recognition epitope is determined through indirect ELISA and Western blotting tests, so that screening is carried out step by step until the shortest linear epitope recognized by the monoclonal antibody is determined, and the amino acid sequence of the shortest linear epitope is shown as SEQ ID No.10. Therefore, the monoclonal antibody and the antigen epitope of the monoclonal antibody can be applied to research on structures and functions of GAstV capsid proteins and preparation of serological diagnostic tools and epitope vaccines.

The invention relates to the technical field of biology, in particular to a variant AAV capsid protein and a recombinant adeno-associated virus rAAV. The variant AAV capsid protein provided by the invention comprises: 1) an amino acid sequence having at least 80%, 83%, 85%, 88%, 90%, 93%, 95% and 98% of identity with an amino acid sequence as shown in SEQ ID NO.1; and 2) an amino acid sequence shown as SEQ ID NO.1, wherein the amino acid sequence comprises amino acid substituted A587D, Q588G, A589N and Q590P, and an amino acid sequence TLAVPFK is inserted after the 588th site and before the 589th site; related products constructed by the variant AAV capsid protein can be used for cross-blood-brain-barrier efficient transduction to realize drug delivery to a central nervous system to treat related diseases of the central nervous system, and meanwhile, the related products have the characteristic of remarkably reducing liver transduction.

The invention relates to a porcine reproductive and respiratory syndrome virus (PRRSV) dimer nucleocapsid (N) protein (PRRSV N-dimer) and application thereof. The PRRSV N-dimer is formed by connecting N protein amino acid sequences in series through (GGGGS) 3linker. According to the invention, a GGGGS 3linker nucleotide sequence is connected in series with a PRRSV NADC30-like strain N protein gene sequence to construct a prokaryotic expression vector, and a recombinant PRRSV N-dimer is expressed in escherichia coli. Through SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and Western-blot detection, the recombinant PRRSV N-dimer obtains soluble expression and can be subjected to a specific reaction with PRRSV positive serum; through detection by a freeze thawing method and an ELISA (Enzyme-Linked Immunosorbent Assay) method, compared with a PRRSV monomer N protein (PRRSV N-monomer), the PRRSV N-dimer protein can be stably preserved, and the immunoreactivity is better. A PRRSV antibody indirect enzyme-linked immunosorbent assay (iELISA) detection method is established on the basis of a PRRSV N-dimer, and can be applied to diagnosis and monitoring of PRRSV infection.

Disclosed are a preparation method and application of a single-chain antibody fragment targeting SARS-COV-2 nucleocapsid protein, aiming to provide a single-chain antibody fragment with high affinity and strong specificity. The present disclosure obtains three single-chain antibody fragments targeting SARS-COV-2 nucleocapsid protein through construction of a nanobody phage library and phage screening technology. The single-chain antibody fragment consists of a heavy chain variable region, a 15aa connecting peptide, and a light chain variable region connected in sequence, and has an amino acid sequence shown in SEQ ID NO: 1-SEQ ID NO: 3. The single-chain antibody fragment in the present disclosure has excellent binding performance and specificity with the SARS-COV-2 nucleocapsid protein, can be applied to a variety of immunoassay platforms of the SARS-COV-2, and has broad application prospects in the field of SARS-COV-2 detection.