Novel recombinant feline interferon carrying N-glycans with reduced allergy risk produced by a transgenic silkworm system.

ABSTRACT: BACKGROUND:The generation of recombinant proteins for commercialisation must be cost-effective. Despite the cost-effective production of recombinant feline interferon (rFeIFN) by a baculovirus expression system, this rFeIFN carries insect-type N-glycans, with core ? 1,3 fucosyl residues that act as potential allergens. An alternative method of production may yield recombinant glycoproteins with reduced antigenicity. RESULTS:A cDNA clone encoding the fifteenth subtype of FeIFN-? (FeIFN-?15) was isolated from a Japanese domestic cat. This clone encoded a protein of 189 amino acids with a molecular mass of 21.1 kDa. The rFeIFN-?15 was expressed using a transgenic silkworm system, which was expected to yield an N-glycan structure with reduced antigenicity compared with the protein produced by the baculovirus system. The resulting rFeIFN-?15 accumulated in the sericin layer of silk fibres and was easily extracted and purified by column chromatography. The N-terminal amino acid sequence of purified rFeIFN-?15 was identical to the mature form of natural sequence. Moreover, its N-glycans did not include detectable core ? 1,3 fucosyl residues. Its anti-vesicular stomatitis virus activity (2.6?×?108 units/mg protein) was comparable to that of the baculovirus-expressed rFeIFN. CONCLUSIONS:The lower allergy risk of rFeIFN produced by the transgenic silkworm system than by the baculovirus expression system is due to the former lacking core ? 1,3 fucosyl residues in its N-glycans. The rFeIFN-?15 produced by the transgenic silkworm system may be a prospective candidate for the next generation of rFeIFN in veterinary medicine.

SUBMITTER: Minagawa S 

PROVIDER: S-EPMC6119277 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature