Abstract

Because of the broad neutralization and in vivo protection across influenza A and influenza B virus strains, monoclonal antibody CR9114 is widely used in influenza virus research as a positive control in many experiments. To produce amounts sufficient for the demand requires regular transient transfections, resulting in varying yield as well as differing batch to batch quality. Here, we report the development of a serum-free CHO DG44 cell line, stably producing a CR9114-like antibody with a potential to become a useful influenza virus research tool.

Highlights

  • Monoclonal antibody CR9114 was first reported by Dreyfus et al in 2012 in a study describing broadly neutralizing human antibodies against influenza B virus strains [1]

  • Chinese hamster ovary (CHO) DG44 cells were cultivated in Dulbecco’s Modified Eagle’s Medium (DMEM)/Ham’s F12 medium supplemented with 10% FBS for 15 passages before the start of the adaptation to serum-free conditions

  • As the FBS concentration was lowered to 2%, the DMEM/Ham’s F12 medium was supplemented as described above and in parallel ProCHO5, CD DG44, OptiCHO, and CHO-S-SFM II media were tested (Fig 1)

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Summary

Introduction

Monoclonal antibody (mAb) CR9114 was first reported by Dreyfus et al in 2012 in a study describing broadly neutralizing human antibodies against influenza B virus strains [1]. What made this antibody so unique is its ability to bind to group 1 (H1, H2, H5, H6, H8, H9, H11, H12, H13, H16, HL17, HL18) and group 2 (H3, H4, H7, H10, H14, H15) influenza A virus hemagglutinins (HA) in addition to influenza B HA by interacting with a highly conserved epitope in the stalk region of HA.

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