Host cell protein quantification by Fourier transform mid infrared spectroscopy (FT-MIR)

Area of Science:

• Biotechnology
• Process Development
• Analytical Chemistry

Background:

• Current methods for monitoring host cell proteins (HCPs) in bioprocessing, such as ELISA, are time-consuming, expensive, and prone to dilution errors at higher concentrations.
• Enhanced monitoring techniques are crucial for efficient process development in upstream and downstream biopharmaceutical manufacturing.

Purpose of the Study:

• To evaluate the suitability of attenuated total reflection (ATR) spectroscopy as a rapid and cost-effective method for quantifying HCPs in monoclonal antibody production.
• To assess ATR spectroscopy's performance in monitoring higher HCP concentrations encountered during process development, potentially eliminating the need for sample dilution.

Main Methods:

• ATR spectroscopy was employed to analyze clarified cell culture fluid from Chinese hamster ovary (CHO) cells producing monoclonal antibodies.
• Samples were treated with different polyelectrolytes for clarification, and multivariate data analysis was performed in the mid-infrared range.
• HCP quantification using ATR spectroscopy was compared against results obtained from traditional ELISA assays.

Main Results:

• ATR spectroscopy demonstrated good agreement with ELISA results for HCP quantification.
• The method proved effective in quantifying HCP titers ranging from approximately 20,000 to 200,000 ng/mL.
• This indicates ATR spectroscopy's suitability for monitoring process development steps with elevated HCP levels.

Conclusions:

• ATR spectroscopy is a viable and advantageous technique for HCP quantification in bioprocess development.
• It offers a non-time-consuming and non-expensive alternative to ELISA, particularly for samples with higher HCP concentrations, by avoiding dilution errors.