The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A
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 ce...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2012-01-01
|
| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1100/2012/263737 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850228151566729216 |
|---|---|
| author | Rita Lasickienė Alma Gedvilaite Milda Norkiene Vaida Simanaviciene Indre Sezaite Dovile Dekaminaviciute Evelina Shikova Aurelija Zvirbliene |
| author_facet | Rita Lasickienė Alma Gedvilaite Milda Norkiene Vaida Simanaviciene Indre Sezaite Dovile Dekaminaviciute Evelina Shikova Aurelija Zvirbliene |
| author_sort | Rita Lasickienė |
| collection | DOAJ |
| description | 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 p16INK4A—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 p16INK4A 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 p16INK4A protein in malignant cervical tissue. Spleen cells of the immunized mice were used to generate monoclonal antibodies against p16INK4A 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. |
| format | Article |
| id | doaj-art-22ca6c264b7c444ea025e34733afd09e |
| institution | OA Journals |
| issn | 1537-744X |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-22ca6c264b7c444ea025e34733afd09e2025-08-20T02:04:37ZengWileyThe Scientific World Journal1537-744X2012-01-01201210.1100/2012/263737263737The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4ARita Lasickienė0Alma Gedvilaite1Milda Norkiene2Vaida Simanaviciene3Indre Sezaite4Dovile Dekaminaviciute5Evelina Shikova6Aurelija Zvirbliene7Institute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaInstitute of Experimental Morphology, Pathology and Anthropology with Muzeum, Bulgarian Academy of Sciences, Acad. G. Bonchev Street, Building 25, 1113 Sofia, BulgariaInstitute of Biotechnology, Vilnius University, Graiciuno 8, 02241 Vilnius, LithuaniaProtein 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 p16INK4A—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 p16INK4A 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 p16INK4A protein in malignant cervical tissue. Spleen cells of the immunized mice were used to generate monoclonal antibodies against p16INK4A 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.http://dx.doi.org/10.1100/2012/263737 |
| spellingShingle | Rita Lasickienė Alma Gedvilaite Milda Norkiene Vaida Simanaviciene Indre Sezaite Dovile Dekaminaviciute Evelina Shikova Aurelija Zvirbliene The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A The Scientific World Journal |
| title | The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A |
| title_full | The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A |
| title_fullStr | The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A |
| title_full_unstemmed | The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A |
| title_short | The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A |
| title_sort | use of recombinant pseudotype virus like particles harbouring inserted target antigen to generate antibodies against cellular marker p16ink4a |
| url | http://dx.doi.org/10.1100/2012/263737 |
| work_keys_str_mv | AT ritalasickiene theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT almagedvilaite theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT mildanorkiene theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT vaidasimanaviciene theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT indresezaite theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT doviledekaminaviciute theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT evelinashikova theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT aurelijazvirbliene theuseofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT ritalasickiene useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT almagedvilaite useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT mildanorkiene useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT vaidasimanaviciene useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT indresezaite useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT doviledekaminaviciute useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT evelinashikova useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a AT aurelijazvirbliene useofrecombinantpseudotypeviruslikeparticlesharbouringinsertedtargetantigentogenerateantibodiesagainstcellularmarkerp16ink4a |