Production of immunogenic human papillomavirus-16 major capsid protein derived virus like particles.
OBJECTIVE
Recombinant DNA technology allows expression of the human papillomavirus (HPV) major capsid protein (L1) in heterologous expression systems and the recombinant protein self assembles to virus-like particles (VLP). We took up this study to produce recombinant HPV-16 L1 in yeast, establish the process of recombinant L1 derived VLP preparation and develop an ELISA using VLP as the antigen for serological evaluation of anti HPV-16 L1 antibody status.


METHODS
Complete HPV-16 L1 was amplified from genomic DNA of an esophageal cancer biopsy, cloned and the protein was expressed in a galactose-inducible Saccharomyces cerevisiae expression system. Self assembled VLP was purified by a two-step density gradient centrifugation process and the VLP preparation used to test its suitability in developing an ELISA.


RESULTS
The recombinant protein was predominantly a ~55 KD species with distinct immunoreactivity and formed VLP as confirmed by electron microscopy. An ELISA using the VLP showed its efficacy in appropriate immunoreactivity to serum/plasma IgG.


CONCLUSIONS
Recombinant HPV-16 capsid protein derived VLP was produced and the VLP antigen based ELISA can be used to probe serological association of HPV with different clinical conditions. The VLP technology can be improved further and harnessed for future vaccine development efforts in the country.
Patel
Mira C
MC
Virology Laboratory, Advanced Centre for Treatment, Research & Education in Cancer (ACTREC) Tata Memorial Centre, Navi, Mumbai.
Patkar
Ketaki K
KK
Basu
Atanu
A
Mohandas
K M
KM
Mukhopadhyaya
Robin
R
D000914:Antibodies, Viral / Q000032:analysis; D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology* / Q000648:ultrastructure; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16