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Tytuł:
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Quantification of prefusion conformation for HIV vaccine using size-exclusion chromatography.
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Autorzy:
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Gollapudi D; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA.
Shadrick W; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA.
Yang Y; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA.
Gowetski DB; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA.
Gall J; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA.
Paula Lei Q; Vaccine Production Program, VRC, NIAID, NIH, 9 W. Watkins Mill Road, Gaithersburg, MD, USA. Electronic address: .
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Źródło:
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Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2022 Jun 30; Vol. 1201-1202, pp. 123296. Date of Electronic Publication: 2022 May 13.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: Amsterdam ; New York : Elsevier, c2002-
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MeSH Terms:
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AIDS Vaccines*/chemistry
HIV-1*
Antibodies, Neutralizing ; Chromatography, Gel ; HIV Antibodies ; Protein Conformation ; env Gene Products, Human Immunodeficiency Virus/metabolism
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Contributed Indexing:
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Keywords: CAP256V2LS; HIV; Meso-scale diagnostics (MSD); Prefusion conformation; Size-exclusion chromatography (SEC); Trimer vaccine (T6931)
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Substance Nomenclature:
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0 (AIDS Vaccines)
0 (Antibodies, Neutralizing)
0 (HIV Antibodies)
0 (env Gene Products, Human Immunodeficiency Virus)
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Entry Date(s):
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Date Created: 20220522 Date Completed: 20220608 Latest Revision: 20220608
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Update Code:
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20240105
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DOI:
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10.1016/j.jchromb.2022.123296
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PMID:
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35598458
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A closed prefusion conformation or an open (non-prefusion) conformational state of a protein vaccine candidate molecule can determine if it effectively elucidates a desired immunity. A quick and reliable method to monitor conformational state is important during vaccine development. In addition to our existing immunoassays, we have developed a unique physicochemical approach using size-exclusion chromatography to assess binding between antibody and the structurally desired antigen protein. Through the bound monoclonal antibody protein vaccine peak shift in the size-exclusion chromatography profile, this method determines the percent closed (prefusion) conformation present in a sample. Since only the closed prefusion conformation binds to the specific antibody, the population of the closed versus the open conformation of the vaccine molecule can be monitored without the need for a reference calibrator. This new method can be applied broadly to vaccine development, as well as for antibody selection during antibody drug discovery. The mAb CAP256V2LS (250 µg/mL) specific to prefusion conformation was mixed with HIV trimer (250 µg/mL) at 2:1 volume ratio, incubated at 37 °C for 30 mins and injected onto HPLC column. The percent of non-prefusion conformation was calculated based on ratio of peak area of unbound trimer and total area of control trimer sample (without mAb).
(Published by Elsevier B.V.)
