The present invention belongs to the field of compliance markers and marker vaccines which allow for the differentiation between infected and vaccinated individuals. In particular, it relates to a method of determining whether an individual has received an immunogenic composition comprising a recombinant protein produced by a baculovirus expression system in cultured insect cells.

Reagents and methods are provided that allow for an improved expression of a recombinant protein. More specifically, the introduction of recombinant DNA elements into a host cell allows for the increased expression of a recombinant protein, an improvement of the correct folding of said protein and an increase in cell viability and proliferation of the host cell, These recombinant DNA elements can be introduced into host cells, for example, via a recombinant baculovirus, which has incorporated said elements. The recombinant DNA elements include nucleic acids encoding transcriptional regulators, such as IE-0 and IE-1, transcriptional enhancer elements, such as the homologous region (hr) and promoters.

A recombinant baculovirus having a) a nucleotide sequence encoding a fusion protein, wherein said fusion protein includes an antigen fused to a baculovirus capsid peptide operatively linked to a first promoter and b) a nucleotide sequence encoding a lipid viral envelope protein bound to a second promoter.

A baculovirus vector and a use thereof in the preparation of a recombinant adeno-associated virus (rAAV) in an insect cell are provided. The baculovirus vector includes an exogenous gene expression cassette and a stable sequence. The stable sequence is located at a site 5 kb or less from the exogenous gene expression cassette, and the stable sequence is a conserved noncoding element (CNE) sequence or a nucleocapsid assembly-essential element (NAE) sequence. When an insect cell is infected with a recombinant baculovirus (rBV) constructed in this way, after multiple continuous passages, production levels of the rBV and the rAAV still remain relatively stable.

A high-volume gene therapy vector manufacturing process which produces a recombinant gene therapy vector which is able to transform host cells even when they are not dividing.

A precision recombinant adeno-associated virus (pciAAV) vector and the use thereof in gene therapy, gene editing, and gene regulation. The pciAAV vector is obtained by packaging an unpackaged pciAAV genome, said pciAAV genome containing the following, in sequence: (a) a modified ITR, which lacks a D element and a trs sequence; (b) a gene of interest or a protection sequence; (c) a complete ITR; (d) a gene of interest or a protection sequence; and (e) a modified ITR, which lacks a D element and a trs sequence; wherein at least one of the segments (b) and (d) contains a gene of interest. In the present invention, the level of impure DNA in the AAV vector is greatly reduced, gene expression efficiency is improved, random integration in the AAV gene vector is reduced, and the risk of gene mutation is reduced by the precision recombinant adeno-associated virus (pciAAV) vector.

A baculovirus vector and a use thereof in the preparation of a recombinant adeno-associated virus (rAAV) in an insect cell are provided. The baculovirus vector includes an exogenous gene expression cassette and a stable sequence. The stable sequence is located at a site 5 kb or less from the exogenous gene expression cassette, and the stable sequence is a conserved noncoding element (CNE) sequence or a nucleocapsid assembly-essential element (NAE) sequence. When an insect cell is infected with a recombinant baculovirus (rBV) constructed in this way, after multiple continuous passages, production levels of the rBV and the rAAV still remain relatively stable.