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Titer measurement of HIV-1 envelope trimeric glycoprotein in cell culture media by a new tandem ion exchange and size exclusion chromatography (IEC-SEC) method.

ABSTRACT: An efficient and specific liquid chromatography (LC)-based assay was developed to monitor the production of recombinant HIV-1 trimeric envelope glycoprotein (HIV Env trimer), a candidate vaccine for HIV-1 infection, in cell culture media to support scale-up process development. In this method, titer measurement was achieved by coupling a weak anion exchange chromatography (IEC) column with a size exclusion chromatography (SEC) column. This assay was specific, accurate, precise, and has been qualified for its intended purpose, with a limit of quantification (LOQ) of 10 µg/mL. This tandem separation strategy offered a reliable and timely analytical support to directly monitor the titer of HIV Env trimer during cell growth, without any extra sample purification steps.

SUBMITTER: Yang G

PROVIDER: S-EPMC8130148 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Titer measurement of HIV-1 envelope trimeric glycoprotein in cell culture media by a new tandem ion exchange and size exclusion chromatography (IEC-SEC) method.

Yang Gengcheng G Schneck Nicole A NA Barefoot Nathan N Yang Yanhong Y Gollapudi Deepika D Arnold Frank J FJ Cooper Jonathan W JW Lei Q Paula QP

Vaccine 20190503 24

An efficient and specific liquid chromatography (LC)-based assay was developed to monitor the production of recombinant HIV-1 trimeric envelope glycoprotein (HIV Env trimer), a candidate vaccine for HIV-1 infection, in cell culture media to support scale-up process development. In this method, titer measurement was achieved by coupling a weak anion exchange chromatography (IEC) column with a size exclusion chromatography (SEC) column. This assay was specific, accurate, precise, and has been qual ...[more]

PMID: 31060952

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