Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Fabrication and characterization of an HPV virus‐like particle (VLP) D‐MAP using centrifugal casting. A) Schematic diagram of the centrifugal casting process for HPV D‐MAP. B) Scanning electron microscopy (SEM) image of the microneedle morphology. C) Bright field microscopic image of HPV D‐MAP. D) Ex vivo evaluation of insertion and delivery performance of 16 V D‐MAP on mouse buccal mucosa. (Top left) Bright‐field microscopy image illustrating microchannels created by trypan blue‐loaded microneedles. (Top right) Bright‐field microscopy image following the application of rhodamine B‐loaded microneedles. (Bottom left) Fluorescence microscopy image demonstrating localized fluorescence at the insertion sites. (Bottom right) Merged bright‐field and fluorescence microscopy image confirming precise microneedle placement and delivery. Microneedles were applied for 20 min prior to tissue section preparation and imaging. E) Antigen content in HPV D‐MAP with ratios of 1:0, 1:20, and 1:50 of VLP to trehalose. F) Comparative analysis of HPV16 L1 VLP by protein electrophoresis, before and after HPV D‐MAP production. G) Dynamic light scattering (DLS) analysis demonstrating particle size distribution. H) Transmission electron microscopy (TEM) image confirming the structural integrity of VLPs encapsulated within 16V D‐MAP. I) Delivery efficiency of 16 V D‐MAP at various application intervals. J) Scanning electron microscopy (SEM) images illustrating the structural changes of microneedles before insertion and after insertion into mouse buccal mucosa at specific time intervals (10 s, 1, 5, 10, 20, and 60 min).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Virus, Electron Microscopy, Ex Vivo, Microscopy, Fluorescence, Imaging, Protein Electrophoresis, Transmission Assay

Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Preformulation assessments for optimal stabilizer composition and long‐term stability evaluation in HPV D‐MAP. The characteristics of HPV16 L1 VLP were evaluated in HPV D‐MAP formulations with ratios of 1:0, 1:20, and 1:50 of VLP to trehalose using A) protein electrophoresis, B) ELISA, C) TEM, and D,E) DLS analysis before and after 7 days under accelerated storage conditions (40 ± 2 °C/75 ± 5% RH). These experiments were conducted with n = 3 per group (A–E). The DLS graphs (D–E) represent mean values including hydrodynamic diameter and PDI (mean±SD). The long‐term stability of HPV D‐MAP, formulated with a 1:50 ratio of VLP to trehalose, was assessed under F) refrigeration (5 ± 3 °C) and G) room temperature (25 ± 2 °C / 60 ± 5% RH), with n = 5 per group (F–G).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Protein Electrophoresis, Enzyme-linked Immunosorbent Assay

Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Delivery of HPV16 VLPs to immune cells via microneedles through buccal tissue. A) Alexa Fluor 647 conjugated HPV16 VLP microneedles (AF647 16 V D‐MAP) were administered to the buccal mucosa of mice for 20 min. Bioluminescence imaging was performed 1 h post‐administration (n = 5 per group). Radiant efficiency was measured in units of (p/s/cm 2 /sr)/(µW/cm 2 ), and the color scale ranged from 1.03 × 10⁸ to 3.45 × 10⁸. B) Quantitative analysis of fluorescence imaging data from the buccal areas of MN Mock and AF647 16 V D‐MAP treated mice. C) Fluorescence imaging of cervical lymph nodes, and D) quantification of antigen delivery to cervical lymph nodes 24 h after microneedle administration. E) Flow cytometric analysis of HPV16 VLP‐specific B cells in the cervical lymph nodes. F) Statistical analysis of HPV16 VLP‐specific cells in cervical lymph nodes. Data are presented as means ± SEM. ns (not significant),p > 0.05; * p < 0.05; ** p ≤ 0.01; **** p ≤ 0.0001 (unpaired t ‐test).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Imaging, Fluorescence

Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Plasma cell differentiation and neutralizing antibody responses induced by 16 V D‐MAP immunization. A) Serum levels of HPV16 VLP‐specific IgG, IgM, and IgA in IM PBS, IM Gardasil, MN Mock, and 16 V D‐MAP groups, detected by ELISA 14 days after the first and third immunization (n = 5 per group). B) Serum neutralization titers against HPV16 PsV were measured at pre‐immunization, 14 days after the first immunization (n = 5 per group), and 14 days after the third immunization (n = 10 per group). C) Enzyme‐linked immunospot (ELISPOT) assay detecting HPV16 VLP‐specific IgG‐, IgM‐, and IgA‐secreting plasma cells in bone marrow (BM) 14 days after the third immunization (n = 5 per group). D) Quantification of IgG‐, IgM‐, and IgA‐secreting plasma cells based on ELISPOT data. Data are presented as means ± SEM. Statistical significance: ns (not significant), p > 0.05; * p < 0.05; * * p ≤ 0.01; ***p ≤ 0.001; * *** p ≤ 0.0001 (unpaired t ‐test).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Clinical Proteomics, Cell Differentiation, Enzyme-linked Immunosorbent Assay, Neutralization, Enzyme-linked Immunospot

Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Protection of mice from buccal and genital challenge with HPV16 PsV by IM PBS, IM Gardasil, MN Mock, or 16 V D‐MAP immunization. A) Bioluminescence imaging of buccal and vaginal mucosal areas in vaccinated mice (n = 5 per group) challenged with HPV16 PsV 14 days after the third immunization. Luminescence scales: buccal, 300–3900; vaginal, 1000–46000. B) Quantitative analysis of bioluminescence imaging data from buccal and vaginal areas. C) Bioluminescence imaging of naïve mice passively transferred with antisera and challenged with buccal HPV16 PsV. Luminescence scales: 200–900. D) Quantitative analysis of HPV16 bioluminescence imaging data from buccal areas. Data are presented as means ± SEM. Statistical significance: ns (not significant), p > 0.05; * p < 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001 (unpaired t ‐test).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Imaging

Journal: Advanced Healthcare Materials

Article Title: Dissolving Microneedle for Maintaining the Integrity of HPV Virus‐Like Particles Enabling Durable Sterile Protection Across Various Mucosal Tissues

doi: 10.1002/adhm.202500963

Figure Lengend Snippet: Long‐term immune response and protection against HPV16 VLP 6 months after 3rd immunization. A) Serum levels of HPV16 VLP‐specific IgG, IgM, and IgA in PBS, IM Gardasil, MN Mock, and 16 V D‐MAP groups, detected by ELISA 6 months after the third immunization (n = 10 per group). B) Serum neutralization titers against HPV16 PsV, measured 6 months after the third immunization (n = 10 per group). C) ELISPOT assay detecting HPV16 VLP‐specific IgG‐, IgM‐, and IgA‐secreting plasma cells in BM 6 months after the third immunization (n = 8 per group). D) Quantification of IgG‐, IgM‐, and IgA‐secreting plasma cells based on ELISPOT data. E) Bioluminescence imaging of buccal and vaginal mucosal areas in vaccinated mice (n = 5 per group) challenged with HPV16 PsV 6 months after the third immunization. F) Quantitative analysis of bioluminescence imaging data from buccal and vaginal areas. Data are presented as means ± SEM. Statistical significance: ns (not significant), p > 0.05; * p < 0.05; * * p ≤ 0.01; *** p ≤ 0.001 (unpaired t ‐test).

Article Snippet: A Maxisorp 96‐well microtiter plate (Thermo Scientific) was coated overnight at 4 °C with a mouse monoclonal antibody specific to HPV16 L1 (CamVir1, Novus Biologicals, Centennial, CO, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Neutralization, Enzyme-linked Immunospot, Clinical Proteomics, Imaging

Journal: Nature reviews. Cancer

Article Title: Opportunities and challenges for human papillomavirus vaccination in cancer

doi: 10.1038/nrc.2018.13

Figure Lengend Snippet: HPV vaccines targeting capsid antigens that are licensed, in or advancing towards clinical trial

Article Snippet: RG1-VLP , HPV-16 L1-L2 (17–36) VLP , BEVS , Aluminium hydroxide , cGMP production , R. Kirnbauer, NCI 130 , Pathovax LLC.

Techniques: Vaccines, Adjuvant

Journal: Infectious Agents and Cancer

Article Title: Serum antibody response to Human papillomavirus (HPV) infections detected by a novel ELISA technique based on denatured recombinant HPV16 L1, L2, E4, E6 and E7 proteins

doi: 10.1186/1750-9378-1-6

Figure Lengend Snippet: Analysis of purified proteins by SDS-PAGE . The purified HPV16 proteins E4, E6, E7, L1 and L2 were run on a polyacrilamide gel electrophoresis and stained by Coomassie blue. Each protein is indicated on the top of the corresponding lane. The weight of the molecular mass markers (lane M) is indicated on the left of the figure.

Article Snippet: The HPV- specific hyperimmune sera to L1, L2, E6, E7 and E4, the commercial anti-Histidine mAb (clone HIS-1, Sigma-Aldrich), anti- HPV16 L1 mAb (clone CamVir-1, Cymbus Biotechnology) and the anti-HPV16E7 mAb (clone 8C9, Zymed Laboratories INC) were used as controls.

Techniques: Purification, SDS Page, Nucleic Acid Electrophoresis, Staining

Journal: Infectious Agents and Cancer

Article Title: Serum antibody response to Human papillomavirus (HPV) infections detected by a novel ELISA technique based on denatured recombinant HPV16 L1, L2, E4, E6 and E7 proteins

doi: 10.1186/1750-9378-1-6

Figure Lengend Snippet: Comparison of the percentages of sero-reactivity . The percentages of sera reacting to the HPV16 L1, L2, E4, E6 and E7 proteins are shown; the percentages of the HPV16 serum group are in white bars and those the other HPVs serum group are in black bars. The percentages of negative sera in the two groups (Neg) are also given.

Article Snippet: The HPV- specific hyperimmune sera to L1, L2, E6, E7 and E4, the commercial anti-Histidine mAb (clone HIS-1, Sigma-Aldrich), anti- HPV16 L1 mAb (clone CamVir-1, Cymbus Biotechnology) and the anti-HPV16E7 mAb (clone 8C9, Zymed Laboratories INC) were used as controls.

Techniques: Comparison

Journal: Infectious Agents and Cancer

Article Title: Serum antibody response to Human papillomavirus (HPV) infections detected by a novel ELISA technique based on denatured recombinant HPV16 L1, L2, E4, E6 and E7 proteins

doi: 10.1186/1750-9378-1-6

Figure Lengend Snippet: HPV primers used in PCR amplification of the HPV 16 genes.

Article Snippet: The HPV- specific hyperimmune sera to L1, L2, E6, E7 and E4, the commercial anti-Histidine mAb (clone HIS-1, Sigma-Aldrich), anti- HPV16 L1 mAb (clone CamVir-1, Cymbus Biotechnology) and the anti-HPV16E7 mAb (clone 8C9, Zymed Laboratories INC) were used as controls.

Techniques: Amplification

Journal: Vaccines

Article Title: Human Papillomavirus-Related Cancer Vaccine Strategies

doi: 10.3390/vaccines12111291

Figure Lengend Snippet: Prophylactic HPV vaccines that have been approved for marketing.

Article Snippet: , VB10.16 , Phase II , L1 proteins of HPV 16 and 18 , VBI Vaccines, Inc., Cambridge, MA, USA.

Techniques: Vaccines, Produced, Expressing, Adjuvant, Injection, Infection

Journal: Vaccines

Article Title: Human Papillomavirus-Related Cancer Vaccine Strategies

doi: 10.3390/vaccines12111291

Figure Lengend Snippet: Globally in-clinical-stage prophylactic and therapeutic HPV vaccines.

Article Snippet: , VB10.16 , Phase II , L1 proteins of HPV 16 and 18 , VBI Vaccines, Inc., Cambridge, MA, USA.

Techniques: Vaccines, Clinical Proteomics