Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types
ABSTRACT Two multiplex immunoassays are routinely used to assess antibody responses in clinical trials of the 9-valent human papillomavirus (9vHPV) vaccine. The HPV6/11/16/18/31/33/45/52/58 competitive Luminex immunoassay (HPV-9 cLIA) and HPV6/11/16/18/31/33/45/52/58 total immunoglobulin G Luminex i...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-04-01
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| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msphere.00962-21 |
| _version_ | 1827963938811150336 |
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| author | Katrina M. Nolan Brent Seaton Joseph Antonello Yuhua Zhang Lauren Cook Krystyn Delfino Leonard J. Rubinstein Kyeongmi Cheon Thomas Group Alain Luxembourg Sheri Dubey |
| author_facet | Katrina M. Nolan Brent Seaton Joseph Antonello Yuhua Zhang Lauren Cook Krystyn Delfino Leonard J. Rubinstein Kyeongmi Cheon Thomas Group Alain Luxembourg Sheri Dubey |
| author_sort | Katrina M. Nolan |
| collection | DOAJ |
| description | ABSTRACT Two multiplex immunoassays are routinely used to assess antibody responses in clinical trials of the 9-valent human papillomavirus (9vHPV) vaccine. The HPV6/11/16/18/31/33/45/52/58 competitive Luminex immunoassay (HPV-9 cLIA) and HPV6/11/16/18/31/33/45/52/58 total immunoglobulin G Luminex immunoassay are used for measurements of immunogenicity. Following their initial validation in 2010, both assays were redeveloped, and several parameters were optimized, including the coating concentration of virus-like particles, type of Luminex microspheres, serum sample and reference standard diluent, reference standard starting dilution and titration series, and vendor and concentration of the phycoerythrin-labeled antibodies. Validation studies evaluated the assay performance parameters, including the intra-assay precision (repeatability), intermediate precision, linearity, relative accuracy, and limits of quantitation. In addition, since maintaining a link to the original assays that were used in trials supporting vaccine licensure is critical, the assays were formally bridged to the previous assay versions by using individual patient sera from a 9vHPV vaccine clinical trial (n = 150 day 1 [prevaccination] samples; n = 100 month 7 [1 month post-last vaccine dose] and n = 100 month 36 [30 months post-last vaccine dose; antibody persistence] samples). The results of the validation studies indicate that both optimized assays are accurate, specific, and precise over their respective quantifiable ranges. There was a strong linear association between the new and previous versions of both assays. Assay serostatus cutoffs for the redeveloped assays were established based on the bridging studies and, for the HPV-9 cLIA, further refined, based on additional data from HPV vaccine clinical studies so as to align the seropositivity rates between assay versions. IMPORTANCE Assay modernization is a key aspect of vaccine life cycle management. Thus, new, reoptimized versions of two 9vHPV immunoassays have been developed and validated for use in ongoing and future HPV vaccine clinical trials. These assays are suitable for use in high-throughput testing for HPV antibodies in serum samples. Bridging to previous versions of the assays allows for the continuous monitoring of immune responses across assay versions, including in immunogenicity studies that involve new populations as well as long-term follow-up studies. |
| first_indexed | 2024-04-09T17:07:02Z |
| format | Article |
| id | doaj.art-fc915b5b917b4300a7a5ca8400155d0e |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-04-09T17:07:02Z |
| publishDate | 2023-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-fc915b5b917b4300a7a5ca8400155d0e2023-04-20T13:01:45ZengAmerican Society for MicrobiologymSphere2379-50422023-04-018210.1128/msphere.00962-21Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine TypesKatrina M. Nolan0Brent Seaton1Joseph Antonello2Yuhua Zhang3Lauren Cook4Krystyn Delfino5Leonard J. Rubinstein6Kyeongmi Cheon7Thomas Group8Alain Luxembourg9Sheri Dubey10Merck & Co., Inc., Rahway, New Jersey, USAQ2 Solutions Vaccines, San Juan Capistrano, California, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAMerck & Co., Inc., Rahway, New Jersey, USAABSTRACT Two multiplex immunoassays are routinely used to assess antibody responses in clinical trials of the 9-valent human papillomavirus (9vHPV) vaccine. The HPV6/11/16/18/31/33/45/52/58 competitive Luminex immunoassay (HPV-9 cLIA) and HPV6/11/16/18/31/33/45/52/58 total immunoglobulin G Luminex immunoassay are used for measurements of immunogenicity. Following their initial validation in 2010, both assays were redeveloped, and several parameters were optimized, including the coating concentration of virus-like particles, type of Luminex microspheres, serum sample and reference standard diluent, reference standard starting dilution and titration series, and vendor and concentration of the phycoerythrin-labeled antibodies. Validation studies evaluated the assay performance parameters, including the intra-assay precision (repeatability), intermediate precision, linearity, relative accuracy, and limits of quantitation. In addition, since maintaining a link to the original assays that were used in trials supporting vaccine licensure is critical, the assays were formally bridged to the previous assay versions by using individual patient sera from a 9vHPV vaccine clinical trial (n = 150 day 1 [prevaccination] samples; n = 100 month 7 [1 month post-last vaccine dose] and n = 100 month 36 [30 months post-last vaccine dose; antibody persistence] samples). The results of the validation studies indicate that both optimized assays are accurate, specific, and precise over their respective quantifiable ranges. There was a strong linear association between the new and previous versions of both assays. Assay serostatus cutoffs for the redeveloped assays were established based on the bridging studies and, for the HPV-9 cLIA, further refined, based on additional data from HPV vaccine clinical studies so as to align the seropositivity rates between assay versions. IMPORTANCE Assay modernization is a key aspect of vaccine life cycle management. Thus, new, reoptimized versions of two 9vHPV immunoassays have been developed and validated for use in ongoing and future HPV vaccine clinical trials. These assays are suitable for use in high-throughput testing for HPV antibodies in serum samples. Bridging to previous versions of the assays allows for the continuous monitoring of immune responses across assay versions, including in immunogenicity studies that involve new populations as well as long-term follow-up studies.https://journals.asm.org/doi/10.1128/msphere.00962-219-valent human papillomavirus vaccinecompetitive Luminex immunoassayhuman papillomavirusimmunogenicitytotal immunoglobulin G immunoassay |
| spellingShingle | Katrina M. Nolan Brent Seaton Joseph Antonello Yuhua Zhang Lauren Cook Krystyn Delfino Leonard J. Rubinstein Kyeongmi Cheon Thomas Group Alain Luxembourg Sheri Dubey Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types mSphere 9-valent human papillomavirus vaccine competitive Luminex immunoassay human papillomavirus immunogenicity total immunoglobulin G immunoassay |
| title | Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types |
| title_full | Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types |
| title_fullStr | Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types |
| title_full_unstemmed | Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types |
| title_short | Development and Validation of Two Optimized Multiplexed Serologic Assays for the 9-Valent Human Papillomavirus Vaccine Types |
| title_sort | development and validation of two optimized multiplexed serologic assays for the 9 valent human papillomavirus vaccine types |
| topic | 9-valent human papillomavirus vaccine competitive Luminex immunoassay human papillomavirus immunogenicity total immunoglobulin G immunoassay |
| url | https://journals.asm.org/doi/10.1128/msphere.00962-21 |
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