The present invention provides a method for efficiently manufacturing a non-enveloped virus with high purity without laborious operation by cultivating cells having the ability to produce a non-enveloped virus and bringing the cells and an acidic solution into contact with each other. A non-enveloped virus vector manufactured by the method of the present invention and a composition having the non-enveloped viral vector as an active ingredient are very useful as gene transfer methods in the fields of basic research and clinical application gene therapy.

The present invention provides a method for efficiently manufacturing a non-enveloped virus with high purity without laborious operation by cultivating cells having the ability to produce a non-enveloped virus and bringing the cells and an acidic solution into contact with each other. A non-enveloped virus vector manufactured by the method of the present invention and a composition having the non-enveloped viral vector as an active ingredient are very useful as gene transfer methods in the fields of basic research and clinical application gene therapy.

Described is an adenovirus derived helper virus which may comprise the adenoviral DNA sequences for E2a, S4 (orf6), the VA1 RNA gene, and the parvovirus VP capsid gene unit. Also described is a method of efficiently preparing a recombinant parvovirus (particle) which is based on the use of various adenoviral derived helper viruses/vectors.

Described is an adenovirus derived helper virus which may comprise the adenoviral DNA sequences for E2a, S4 (orf6), the VA1 RNA gene, and the parvovirus VP capsid gene unit. Also described is a method of efficiently preparing a recombinant parvovirus (particle) which is based on the use of various adenoviral derived helper viruses/vectors.

Provided are establishment and suspension acclimation of a Crandell Reese Feline Kidney (CRFK) adherent cell line, and its application. The CRFK cell line, named CRFK-BLA, is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45703; the CRFK cell line was classified and named CRFK cells, and was deposited on Aug. 17, 2023. A serum-free complete suspension culture type CRFK cell line obtained by acclimation on the basis of CRFK-BLA is named CRFK-BLS, and is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45704; the CRFK cell line was classified and named CRFK suspension cells.

Provided are establishment and suspension acclimation of a Crandell Reese Feline Kidney (CRFK) adherent cell line, and its application. The CRFK cell line, named CRFK-BLA, is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45703; the CRFK cell line was classified and named CRFK cells, and was deposited on Aug. 17, 2023. A serum-free complete suspension culture type CRFK cell line obtained by acclimation on the basis of CRFK-BLA is named CRFK-BLS, and is deposited at the China General Microbiological Culture Collection Center (CGMCC), with the address being Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, the deposit number being CGMCC NO: 45704; the CRFK cell line was classified and named CRFK suspension cells.

A heavy-chain variable region sequence of the nanobody CPV-VHH-H1 has the amino acid sequence set forth in SEQ ID NO: 1; and a gene encoding the nanobody CPV-VHH-H1 has the nucleotide sequence set forth in SEQ ID NO: 2. A nanobody immune library of the CPV is constructed by a phage display technology, a specific anti-CPV nanobody CPV-VHH-H1 is obtained by screening, and it is verified by experiments that the nanobody may specifically bind to the CPV. A new nanobody preparation for use in clinical diagnosis and treatment of the CPV can be developed, and a certain theoretical reserve is provided for applying the nanobody to the field of veterinary biological products.

The present invention provides a robust single clone Master Cell Bank (MCB) for an optimized production of H-1 parvovirus (H-1 PV) which is suitable to increase infectious parvovirus production compared to standard producer NB-324K mixed cells.