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Fast, selective and quantitative protein profiling of adenovirus-vector based vaccines by ultra-performance liquid chromatography
Janssen Vaccines & Prevent Pharmaceut & Analyt De, Newtonweg 1, NL-2333 CP Leiden, Netherlands.
Janssen Vaccines & Prevent Pharmaceut & Analyt De, Newtonweg 1, NL-2333 CP Leiden, Netherlands.
Janssen Vaccines & Prevent Pharmaceut & Analyt De, Newtonweg 1, NL-2333 CP Leiden, Netherlands.
Janssen Vaccines & Prevent Pharmaceut & Analyt De, Newtonweg 1, NL-2333 CP Leiden, Netherlands.
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2018 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1581-1582, p. 25-32Article in journal (Refereed) Published
Abstract [en]

A method for the quantitative determination of the protein composition of adenovirus-vector based vaccines was developed. The final method used RP-UPLC with UV absorbance detection, a C4 column (300 angstrom, 1.7 mu m, 2.1 x 150 mm), and a water- acetonitrile (ACN) gradient containing trifluoroacetic acid (TFA) as ion-pairing agent. The chromatographic resolution between the various adenovirus proteins was optimized by studying the effect of the TFA concentration and the column temperature, applying a full factorial design of experiments. A reproducible baseline separation of all relevant adenovirus proteins could be achieved within 17 min run time. Samples containing adenovirus particles could be directly injected into the UPLC system without sample pretreatment. The viruses reproducibly dissociate into proteins in the UPLC system upon contact with the mobile phase containing ACN. The new RP-UPLC method was successfully validated for protein profiling and relative quantification of proteins in vaccine products based on adenovirus vector types 26 and 35. The intermediate precision of the relative peak areas of all proteins was between 1% and 14% RSD, except for the peak assigned to protein V (26% RSD). The method proved to be stability indicating with respect to thermal and oxidation stress of the adenovirus-vector based vaccine and was successfully implemented for the characterization of adenovirus-based products.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE BV , 2018. Vol. 1581-1582, p. 25-32
Keywords [en]
Intact proteins, Adenovirus, Protein profiling, RP-UPLC, Quantification
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-372884DOI: 10.1016/j.chroma.2018.10.045ISI: 000453490100004PubMedID: 30389208OAI: oai:DiVA.org:uu-372884DiVA, id: diva2:1277112
Available from: 2019-01-09 Created: 2019-01-09 Last updated: 2019-10-18Bibliographically approved

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Sänger - van de Griend, Cari

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