Key result
Recombinant human exCAR produced in insect cells yielded 8-10 mg/L with 96.3% purity and retained its ability to bind to the adenovirus type 5 fiber protein knob domain.
Recombinant human exCAR can be produced in large scale in insect cells with high purity and retained binding activity, potentially useful for adenovirus infection therapy.
Scalable exCAR production with retained activity supports further preclinical work; animal data leave clinical translation open.
Coxsackievirus group B and adenovirus receptor (CAR) is a major receptor for the adenovirus groups that has drawn overall attention over the past decade. Although this protein could potentially be used as an agent for the blocking of adenovirus infection, large-scale production of highly purified human CAR in eukaryotic expression system has not been reported. In the present study, we showed the construction of recombinant baculovirus highly-expressing the extracellular domain of human coxsackievirus-adenovirus receptor (exCAR) in High Five insect cells. The recombinant exCAR was recovered from the cell culture medium as a secreted soluble protein and purified by Ni-NTA affinity chromatography. The final yield of recombinant exCAR was about 8-10 mg/l of supernatant with the purity of 96.3%. Binding activity assay showed that the recombinant exCAR exhibited an intact ability of binding to the knob domain of the adenovirus type 5 fiber protein (Ad fiber knob) displayed by T7 phage. These results showed that the recombinant human exCAR produced in insect cells and purified by Ni-NTA chromatography retained its ability to bind to the Ad fiber knob and could potentially be used in therapy of adenovirus infection.
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Zhou et al. (2011) studied Adenovirus infection. Recombinant human exCAR produced in insect cells was evaluated on Yield, purity, and binding activity of recombinant exCAR. Recombinant human exCAR produced in insect cells yielded 8-10 mg/L with 96.3% purity and retained its ability to bind to the adenovirus type 5 fiber protein knob domain.
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