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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Expression and characterization of HPV-16 L1 capsid protein in Pichia pastoris

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Bazan, Silvia Boschi [1, 2] ; Muniz Chaves, Agtha de Alencar [2] ; Aires, Karina Araujo [2] ; Cianciarullo, Aurora Marques [3] ; Garcea, Robert L. [4] ; Ho, Paulo Lee [1, 2]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Quim, Dept Bioquim, BR-01498 Sao Paulo - Brazil
[2] Inst Butantan, Ctr Biotecnol, BR-05503900 Sao Paulo - Brazil
[3] Inst Butantan, Dept Genet, BR-05503900 Sao Paulo - Brazil
[4] Univ Colorado, Dept Mol Cellular & Dev Biol, Boulder, CO 80309 - USA
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: ARCHIVES OF VIROLOGY; v. 154, n. 10, p. 1609-1617, OCT 2009.
Citações Web of Science: 38
Assunto(s):Saccharomyces cerevisiae   Escherichia coli

Human papillomaviruses (HPVs) are responsible for the most common human sexually transmitted viral infections. Infection with high-risk HPVs, particularly HPV16, is associated with the development of cervical cancer. The papillomavirus L1 major capsid protein, the basis of the currently marketed vaccines, self-assembles into virus-like particles (VLPs). Here, we describe the expression, purification and characterization of recombinant HPV16 L1 produced by a methylotrophic yeast. A codon-optimized HPV16 L1 gene was cloned into a non-integrative expression vector under the regulation of a methanol-inducible promoter and used to transform competent Pichia pastoris cells. Purification of L1 protein from yeast extracts was performed using heparin-sepharose chromatography, followed by a disassembly/reassembly step. VLPs could be assembled from the purified L1 protein, as demonstrated by electron microscopy. The display of conformational epitopes on the VLPs surface was confirmed by hemagglutination and hemagglutination inhibition assays and by immuno-electron microscopy. This study has implications for the development of an alternative platform for the production of a papillomavirus vaccine that could be provided by public health programs, especially in resource-poor areas, where there is a great demand for low-cost vaccines. (AU)

Processo FAPESP: 04/11277-4 - Expressao da proteina l1 do capsidio de hpv-16 em leveduras metilotroficas.
Beneficiário:Silvia Boschi Bazan
Linha de fomento: Bolsas no Brasil - Mestrado