title: Solubility as a limiting factor for expression of hepatitis A virus proteins in insect cell-baculovirus system.
abstract: The use of recombinant proteins may represent an alternative model to inactivated vaccines against hepatitis A virus (HAV). The present study aimed to express the VP1 protein of HAV in baculovirus expression vector system (BEVS). The VP1 was expressed intracellularly with molecular mass of 35 kDa. The VP1 was detected both in the soluble fraction and in the insoluble fraction of the lysate. The extracellular expression of VP1 was also attempted, but the protein remained inside the cell. To verify if hydrophobic characteristics would also be present in the HAV structural polyprotein, the expression of P1-2A protein was evaluated. The P1-2A polyprotein remained insoluble in the cellular extract, even in the early infection stages. These results suggest that HAV structural proteins are prone to form insoluble aggregates. The low solubility represents a drawback for production of large amounts of HAV proteins in BEVS.
authors: Fundação Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos, Laboratório de Tecnologia Recombinante, Rio de Janeiro, RJ, Brasil.
authors: Haroldo Cid da
authors: HC
authors: Silva
authors: Fundação Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos, Laboratório de Tecnologia Recombinante, Rio de Janeiro, RJ, Brasil.
authors: Cristiane Pinheiro
authors: CP
authors: Pestana
authors: Fundação Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos, Laboratório de Tecnologia Recombinante, Rio de Janeiro, RJ, Brasil.
authors: Ricardo
authors: R
authors: Galler
authors: Fundação Oswaldo Cruz, Instituto de Tecnologia em Imunobiológicos, Laboratório de Tecnologia Recombinante, Rio de Janeiro, RJ, Brasil.
authors: Marco Alberto
authors: MA
authors: Medeiros
mesh_terms: D016367:Baculoviridae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D030041:Hepatitis A virus / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D014764:Viral Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins