Recombinant viral proteins for use in diagnostic ELISAs to detect virus infection

Abstract

ELISAs provide a valuable tool in the detection and diagnosis of virus infection. The ability to produce recombinant viral proteins will ensure that future ELISAs are safe, specific and rapid. This latter point being the most crucial advantage in that even if a virus cannot be cultured, provided gene sequence is available, it is possible to rapidly respond to emerging viruses and new viral strains of existing pathogens. Indeed, ELISAs based on peptides (corresponding to epitopes) also hold great promise, as in this case no cloning or expression of a recombinant protein is required. Both recombinant protein and peptide based systems lend themselves to large scale production and purification. These approaches can also be used to distinguish recombinant vaccines from parental or wild type viruses.

A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection

“…The low viral yield is a disadvantage of working with cell culture antigens compared with the use of recombinant proteins, especially for the preparation of a high-quality antigen (Nawagitgul et al 2002, Spencer et al 2007). The use of purified virus only for the production of capture antibody reduced the amount of reagent required for the development of the method.…”

“…For the detection of antibodies against PCV2, the systems of antigen production in Escherichia coli (Shang et al 2008, Marcekova et al 2009, Sun et al 2010, Ge et al 2012, insect cells (Nawagitgul et al 2002, Blanchard et al 2003, Liu et al 2004) and insect larvae (Pérez-Martín et al 2008) exhibit more restricted access to techniques when compared with traditional antigen production in cell culture. However, these systems exhibit high purified protein yields (Nawagitgul et al 2002, Spencer et al 2007), which could be used in indirect ELISA methods. Production of pure or partly purified viral antigens with good yield commonly used in these ELI-SA methods is highly dependent on the amount of antigen mass that is produced.…”

Antigenic and immunogenic properties of the canine distemper virus nucleocapsid protein expressed in Escherichia coli employing codon optimized synthetic gene

“…There are many options to express recombinant proteins depending on the specific requirement (Spencer et al 2007), however, Escherichia coli expression dominates the expression systems and remains the first choice for laboratory investigations (Papaneophytou & Kontopidis 2014). The expression in E. coli is relatively simple and a high yield of protein can be produced (LaVallie 2001).…”

Biophysical characterisation of the nucleocapsid protein from a highly pathogenic porcine reproductive and respiratory syndrome virus strain

“…Although vaccines to a number of coronaviruses and arteriviruses exist, the high mutation rates during virus replication leads to new viral threats that are constantly emerging [4]. A more molecular understanding of viral protein function would allow the development of new vaccines, identify targets for targeted antiviral therapy and provide robust diagnostics [5,6].…”