Patent classifications
C12N2710/14143
PREPARATION METHOD AND SYSTEM FOR RECOMBINANT ADENO-ASSOCIATED VIRUS, AND RECOMBINANT BACMID
Provided are a preparation method and system for a recombinant adeno-associated virus (rAAV) and a recombinant bacmid. The method comprises: first reconstructing a recombinant bacmid containing a recombinant baculovirus genome that produces essential functional elements for an rAAV, at least one of the essential functional elements being inserted into the N-terminal or C-terminal of a locus of an essential gene of the recombinant baculovirus genome; and then transfecting the obtained recombinant bacmid containing the recombinant baculovirus genome that produces the rAAV into a host cell line for culturing to prepare an rAAV. Compared with recombinant baculoviruses obtained by conventional Tn7 recombinant preparations of recombinant bacmid, the recombinant baculovirus obtained by inserting a core element containing Cap, Rep and ITR into two sides of a baculovirus essential gene has a more stable rAAV serial passage production level in a cell and has a higher rAAV yield.
VECTORIZED EDITING OF NUCLEIC ACIDS TO CORRECT OVERT MUTATIONS
The disclosure relates to compositions, methods, and processes for the preparation, use, and/or formulation of adeno-associated virus (AAV) particle comprising a viral gene and a capsid, wherein the viral genome comprises at least one Vectorized Editing of Nucleic acids to correct Overt Mutations (VENOM) element.
RECOMBINANT ANTIGEN AND METHOD OF MAKING THE SAME, ISOLATED POLYNUCLEOTIDE OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS (PRRSV) AND IMMUNOGENIC COMPOSITION INCLUDING THE SAME
The present invention relates to a recombinant antigen and an isolated polynucleotide of porcine reproductive and respiratory syndrome virus (PRRSV), a composition including the same and a method of making the same. The recombinant antigen is a chimeric protein of PRRSV dual structural proteins and T-cell epitope. The polynucleotide encodes an amino acid sequence of the recombinant antigen. The recombinant protein expressed by the polynucleotide in an eukaryotic expression system can be beneficial for mass production and purification. An immunogenic composition including the recombinant antigen can promote pro-inflammatory M1-phenotype polarization of porcine alveolar macrophages (PAMs), reduce receptor CD163 expression that is mediated for viral entry and activate T helper (Th1) immune responses, thereby being applied to a vaccine composition against PRRSV.
BACULOVIRUS EXPRESSION SYSTEM
The invention relates to a method for producing a recombinant baculovirus comprising n exogenous genes in an insect cell, by means of homologous recombination of a replication-deficient baculovirus genome and n transfer vectors, each comprising one of the n exogenous genes, n being an integer at least equal to 2.
RECOMBINANT VIRUSES, INSECT CELLS AND THEIR USES IN VIRAL DETECTION AND VACCINATION
Disclosed herein are recombinant viruses and/or insect cells suitable for detecting the infection of a pathogen in a biological sample of a test subject. The information derived from the detection may also be used to render a diagnosis on whether the test subject is infected with the pathogen or not, so that proper course of treatment may be assigned to the subject. Also disclosed herein is a vaccine for the prophylaxis and/or treatment of infection caused by said pathogen.
MACHINE GUIDED DIRECTED EVOLUTION OF RAAV COMBINATORIAL CAPSID LIBRARIES
Provided herein are methods and compositions useful in the screening and production of recombinant AAV (rAAV) that have structural fitness and potency in insect cells and mammalian cells. The disclosure provides a directed evolution system for generating and selecting rAAV variants based on their Kozak sequences. In some embodiments, methods disclosed herein comprise the use of modified Kozak sequences to express AAV2 VP1 proteins in amounts that are useful for producing rAAV particles having high infectivities and/or transduction efficiencies. In some embodiments, methods of producing, and methods of screening, rAAV are provided that comprise the steps of infecting insect cells by administering rAAV to these cells, recovering rAAV-integrated DNA from these insect cells, and transfecting recovered rAAV into mammalian cells. Also provided herein are nucleic acids comprising nucleotide sequences encoding novel Kozak sequences. Further provided herein are mammalian cells, baculovirus particles and insect cells comprising these nucleic acids.
Donor plasmid vectors
Certain donor plasmid vectors such as pFastBac™1 and pFastBac™ Dual lack a cis DNA element upstream of the polh translation start codon (ATG) present in wild type (wt) Autographa californica multiple nucleopolyhedrovirus (AcMNPV), and contain a SV40 pA fragment. When a cis DNA element is inserted upstream of the 50 bp polh promoter and SV40 pA was replaced with a AcMNPV polh pA signal in pFastBac™1 and pFastBac™Dual, certain protein expression levels in High Five™ cells using the Bac-to-Bac® system reached that of the wt AcMNPV.
METHODS AND KITS FOR THE DIAGNOSIS OF CANCER THROUGH DETECTION OF HUMAN ENDOGENOUS RETROVIRUS ENV (HERV-WL)
The present invention relates to methods and kits for diagnosis of cancer in a subject by detecting human endogenous retrovirus env (HERV-WL) polypeptides.
Vaccine compositions having improved stability and immunogenicity
Disclosed herein are nanoparticles suitable for use in vaccines. The nanoparticles present antigens from pathogens surrounded to and associated with a detergent core resulting in enhanced stability and good immunogenicity. Dosages, formulations, and methods for preparing the vaccines and nanoparticles are also disclosed.
Vectors
The present invention relates to a transfer vector for inserting a gene into a genetic locus of a baculovirus sequence. The transfer vector comprises an expression cassette comprising a eukaryotic promoter operably linked to the gene and a bipartite selection cassette. The present invention also relates to methods of using the transfer vector and derived bacmids and baculoviruses.