Identification and Root Cause Analysis of Cell Culture Media Precipitates in the Viral Deactivation Treatment with High-Temperature/Short-Time Method

Abstract

Viral contamination in cell culture media is an important issue in the biopharmaceutical manufacturing industry and may have serious consequences on product quality, efficacy, and safety. High-temperature/short-time (HTST) treatment of cell culture media is one of the proven techniques used in the industry for the prevention and mitigation of media viral contamination. With the HTST method, the formulated media is pasteurized (virus-deactivated) by heating at preset conditions. This paper provides the identification and root-cause study of the media precipitates that adversely affected the HTST process and discusses the possible solutions to mitigate the precipitation problem.

Microspectroscopic investigation of the membrane clogging during the sterile filtration of the growth media for mammalian cell culture

“…The iron might arise from the inorganic compound iron (III) phosphate, which is known for its propensity to precipitate due to its very low aqueous solubility [5]. Iron (III) phosphate also exhibits a distinct P-O stretching band around 1060 cm −1 from the PO 4 3− group [5], which overlaps with the NA bands in this region. Thus the broader and stronger band at ∼1080 cm −1 in Fig.…”

“…4 (red trace) may provide further evidence for the presence of the iron phosphates at the stained area. The iron (III) phosphates might come from the precipitation of two soluble media ingredients: iron chloride and sodium phosphates by means of ion recombination [5], which could occur when the media inside the filter became concentrated due to the membrane clogging. …”

“…The ATR-IR technique has been used extensively in the studies of membrane filters [7][8][9][10] for separation and purifications. The EDS can provide elemental compositions of the samples, which are complementary to the infrared data and aid in the analysis of low-levels of inorganics [4,5].…”

“…In this study, we report the root cause analysis of the clogged sterile filtration based on multiple microspectroscopic techniques [4][5][6], including optical microscopy (OM), fluorescence microscopy (FM), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) microscopy and energy dispersive x-ray spectroscopy (EDS). OM was used for visual inspection and imaging of targeted samples.…”

Treating cell culture media with UV irradiation against adventitious agents: Minimal impact on CHO performance

“…8 Applying HTST can also cause phosphate-and calcium-based precipitates to form, ultimately impacting the operation of the HTST system. 9,10 Alternative to heat treatments, aseptic procedures require all production equipment and components to be either "single-use" disposable or steam-sterilized before use. Cell media are often subjected to sterile filtration with 0.22 mm filters following aseptic steps to further preclude media contamination.…”