Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector.
Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.
Laboratório de Estudos Moleculares e Terapia Experimental, Departamento de Genética, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, PE, Brasil.
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Coimbra
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Gomes
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Campos
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D'arc
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Carvalho
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Mariz
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Jesus
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Stocco
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Beçak
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Freitas
D052685:Alphapapillomavirus / Q000235:genetics / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D002472:Cell Transformation, Viral / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D015967:Gene Expression Regulation, Viral; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D053918:Papillomavirus Vaccines / Q000276:immunology; D010843:Pichia / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000914:Antibodies, Viral; D036022:Capsid Proteins; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines