The use of recombinant pseudotype virus-like particles harbouring inserted target antigen to generate antibodies against cellular marker p16INK4A.
ScientificWorldJournal
; 2012: 263737, 2012.
Article
en En
| MEDLINE
| ID: mdl-22629125
Protein engineering provides an opportunity to generate new immunogens with desired features. Previously, we have demonstrated that hamster polyomavirus major capsid protein VP1-derived virus-like particles (VLPs) are highly immunogenic and can be employed for the insertion of foreign epitopes at certain surface-exposed positions. In the current study, we have designed pseudotype VLPs consisting of an intact VP1 protein and VP2 protein fused with the target antigen--cellular marker p16(INK4A)--at its N terminus. Both proteins coexpressed in yeast were self-assembled to pseudotype VLPs harbouring the inserted antigen on the surface. The pseudotype VLPs were used for generation of antibodies against p16(INK4A) that represents a potential biomarker for cells transformed by high-risk human papillomavirus (HPV). The pseudotype VLPs induced in immunized mice a strong immune response against the target antigen. The antisera raised against pseudotype VLPs showed specific immunostaining of p16(INK4A) protein in malignant cervical tissue. Spleen cells of the immunized mice were used to generate monoclonal antibodies against p16(INK4A) protein. The specificity of antibodies was proven by the immunostaining of HPV-transformed cells. In conclusion, the current study demonstrates the potential of pseudotype VLPs with inserted target antigen as a new type of immunogens to generate antibodies of high diagnostic value.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Autoanticuerpos
/
Virión
/
Proteínas Recombinantes de Fusión
/
Ingeniería de Proteínas
/
Inhibidor p16 de la Quinasa Dependiente de Ciclina
Límite:
Animals
Idioma:
En
Revista:
ScientificWorldJournal
Asunto de la revista:
MEDICINA
Año:
2012
Tipo del documento:
Article
País de afiliación:
Lituania
Pais de publicación:
Estados Unidos