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

Vaccine. 2019 May 27;37(24):3142-3145. doi: 10.1016/j.vaccine.2019.04.089. Epub 2019 May 3.

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.

Keywords: Cell culture; HIV-1; HPLC; Quantification; Tandem chromatography; Titer; Vaccine.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Chromatography, Gel / methods*
  • Chromatography, Ion Exchange / methods*
  • Culture Media / chemistry*
  • Glycoproteins / isolation & purification*
  • HIV-1
  • Protein Multimerization
  • Recombinant Proteins / isolation & purification
  • Reproducibility of Results
  • env Gene Products, Human Immunodeficiency Virus / isolation & purification*

Substances

  • Culture Media
  • Glycoproteins
  • Recombinant Proteins
  • env Gene Products, Human Immunodeficiency Virus