l2 pp7 vlps  (Lucigen Corp)

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: The consensus sequence was derived from an alignment of 15 carcinogenic HPVs, 2 genital HPVs, 4 cutaneous HPVs plus 2 animal papillomaviruses using ClustalW2. Recombinant L2 PP7 VLPs displaying these peptide sequences were constructed. Amino acids are shown in single-letter code, dots indicate that the amino acid is identical to consensus sequence, and amino acid differences from the consensus are shown for each HPV type. A plus (+) symbol indicates that there is no consensus amino acid at this position.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Sequencing, Derivative Assay, Recombinant, Construct

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Shown are A) wild-type PP7 VLPs, B) 16L2 PP7 VLPs, and C) 18L2 PP7 VLPs. VLPs were visualized at a magnification of 40,000×.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques:

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: 500 ng of wild-type PP7 VLPs or L2 PP7 VLPs were used to coat ELISA plates. Binding of a 1∶5,000 dilution of RG-1 was detected using a horseradish peroxidase-conjugated goat anti-mouse IgG secondary antibody followed by development with 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS). Reactivity was determined by measuring the mean optical density (OD) values at 405 nm. Error bars indicate standard error of the mean (SEM) of duplicate wells.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Mice were immunized i.m. twice with 5 µg of PP7 or L2 PP7 VLPs with IFA and their sera collected 2 weeks after the last immunization. Serum anti-L2 IgG titer was determined by end-point dilution ELISA against streptavidin-conjugated HPV L2 peptides (amino acid 14–40) from HPV1 (squares), HPV5 (inverted triangles), HPV6 (upright triangles), HPV16 (white-filled circles), and HPV18 (diamonds). Black-filled circles denote reactivity of PP7 sera with the different five L2 peptides. Each datum point shows antibody titer from each mouse and lines represent the geometric mean titers for each group.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: (A) ELISA plates were coated with the streptavidin-conjugated L2 peptides described in and reacted with a 1∶160 dilution of serum from mice immunized with wild-type PP7 VLPs or L2 PP7 VLPs followed by secondary antibody. Shown are the averages of the optical density (OD 405 ) values of sera from three individual mice in each group. Error bars represent SEM. (B) A summary of these results. “++++” indicates a mean OD 405 >1.2, “+++” indicates a mean OD 405 between 0.8 and 1.2, “++” indicates a mean OD 405 between 0.4 and 0.8, “+” indicates a mean OD 405 between 0.2 and 0.4, and “−” indicates a mean OD 405 below 0.2.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Groups of 5 BALB/c mice were immunized i.m. twice with 5 µg of PP7 VLPs, 16L2 PP7 VLPs or 18L2 PP7 VLPs with IFA. Three weeks after the second immunization, mice were vaginally challenged with 1.3×10 5 (PsV18) or 3.0×10 6 (PsV16) IU of PsV. Forty-eight hours later, luciferin was instilled vaginally and images were taken 3 minutes post-luciferin instillation. Images showing the magnitude of vaginal infection with PsV18 or PsV16 are shown in panels A and B, respectively. The colors reflect the intensity of luciferase expression. Colors are scaled for each image and shown to the right of the image. Quantitative data (shown in each panel) was extracted by drawing equally sized regions of interests surrounding the site of PsV instillation and determining average radiance (p/s/cm 2 /sr) by using Living Image 3.2 software. Background radiance (determined by gating on another region of the mouse) was subtracted from this value. Black-filled circles denote mice immunized with control wild-type PP7 VLPs and white-filled circles denote mice immunized with either 16L2 PP7 VLPs or 18L2 PP7 VLPs. Lines reflect the geometric mean radiance for each group.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Infection, Luciferase, Expressing, Software, Control

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Shown is the reactivity of sera from mice immunized with mixed L2 PP7 VLPs to L2 peptides. Mice were immunized i.m. three times with 10 µg of wild-type PP7 VLPs or 10 µg of mixed L2 PP7 VLPs. Sera were collected 2 weeks after the last immunization and serum anti-L2 IgG titer was determined by end-point dilution ELISA against streptavidin-conjugated HPV L2 peptides (amino acid 14–40) from HPV1, HPV5, HPV6, HPV16, and HPV18. Black-filled circles denote mice immunized with PP7 VLPs and diamonds denote mice immunized with mixed L2 PP7 VLPs.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Groups of 5 Balb/c mice were immunized i.m. three times with 10 µg of PP7 VLPs or mixed L2 PP7 VLPs. Three weeks after the last immunization, mice were vaginally challenged with 1.3×10 5 – 6.5×10 6 IU of PsV5, PsV6, PsV16, PsV18, PsV31, PsV45, PsV52, or PsV58. Forty-eight hours later, luciferin was instilled vaginally and images were taken 3 minutes post-luciferin instillation. Average radiance (p/s/cm 2 /sr) values for each animal were determined as described in . Black-filled circles denote mice immunized with wild-type PP7 VLPs and white-filled circles denote mice immunized with mixed L2 PP7 VLPs.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques:

Journal: PLoS ONE

Article Title: A Pan-HPV Vaccine Based on Bacteriophage PP7 VLPs Displaying Broadly Cross-Neutralizing Epitopes from the HPV Minor Capsid Protein, L2

doi: 10.1371/journal.pone.0023310

Figure Lengend Snippet: Balb/c mice were immunized i.m. as described in . Three weeks after the last immunization, mice were subcutaneously challenged in the belly with 6.0×10 5 IU of PsV5. Three days post-challenge, mice were anesthetized and 0.7 mg luciferin was injected subcutaneously. Images of mice immunized with control PP7 VLPs and mixed L2 PP7 VLPs are shown in panels A and B, respectively. Average radiance values for the two groups are shown in panel C. Black-filled circles denote mice immunized with PP7 VLPs and white-filled circles denote mice immunized with mixed L2 PP7 VLPs.

Article Snippet: L2 PP7 VLPs were made by transforming CSH41F or C41 cells (Lucigen) with plasmids p2P7K32 and pET2P7K32, respectively, containing L2 peptides from different HPV types.

Techniques: Injection, Control