Regeneration of Capto Core 700 resin through high throughput and laboratory scale studies and impact on production of a SARS‐CoV‐2 vaccine candidate

Abstract

During the development of a SARS‐CoV‐2 vaccine candidate, at the height of the COVID‐19 pandemic, raw materials shortages, including chromatography resins, necessitated the determination of a cleaning in place (CIP) strategy for a multimodal core‐shell resin both rapidly and efficiently. Here, the deployment of high throughput (HT) techniques to screen CIP conditions for cleaning Capto Core 700 resin exposed to clarified cell culture harvest (CCCH) of a SARS‐CoV‐2 vaccine candidate produced in Vero adherent cell culture are described. The best performing conditions, comprised of 30% n‐propanol and ≥0.75 N NaOH, were deployed in cycling experiments, completed with miniature chromatography columns, to demonstrate their effectiveness. The success of the CIP strategy was ultimately verified at the laboratory scale. Here, its impact was assessed across the entire purification process which also included an ultrafiltration/diafiltration step. It is shown that the implementation of the CIP strategy enabled the re‐use of the Capto Core 700 resin for up to 10 cycles without any negative impact on the purified product. Hence, the strategic combination of HT and laboratory‐scale experiments can lead rapidly to robust CIP procedures, even for a challenging to clean resin, and thus help to overcome supply shortages.

Understanding the Spike Protein in COVID-19 Vaccine in Recombinant Vesicular Stomatitis Virus (rVSV) Using Automated Capillary Western Blots

“…The downstream purification process for V590 production has been previously described . In brief, the V590 COVID-19 vaccine replaced the G-protein gene in rVSV with the S-protein gene and it was produced in Vero cells in a single-use bioreactor containing Cytodex-1 γ-irradiated microcarriers (Cytiva) and GIBCO VP-SFM media (Thermo Fisher Scientific, Inc.) supplemented with l -glutamine and P-188.…”

“…The downstream purification process for V590 production has been previously described. 28 In brief, the V590 COVID-19 vaccine replaced the G-protein gene in rVSV with the S-protein gene and it was produced in Vero cells in a single-use bioreactor containing Cytodex-1 γ-irradiated microcarriers (Cytiva) and GIBCO VP-SFM media (Thermo Fisher Scientific, Inc.) supplemented with l -glutamine and P-188. Ninety min before harvest, Benzonase endonuclease (MilliporeSigma, MA) and magnesium chloride were added directly to the bioreactor to final concentrations of 60 U/mL and 2 mM, respectively.…”

Adaptation of an rVSV Ebola vaccine purification process for rapid development of a viral vaccine candidate for SARS‐CoV‐2

“…A miniaturized and automated plaque assay was used to determine viral potency measurements, as previously described. [22] Vero cells were seeded into 96-well tissue culture microplates at 40,000 cells per well, and incubated overnight at 37 • C, 5% pCO 2 , >90% relative humidity (rH) until 90% confluence was achieved. The cells were infected by adding the prepared virus samples to each well.…”

Removal of chromatin by salt‐tolerant endonucleases for production of recombinant measles virus

“…This resin material is specifically used for the purification of viruses and bigger particles as they cannot enter the pores 15,16 . At the inside of the pores, positively and hydrophobically active ligands can bind the process related impurities like host cell proteins and DNA 6,17,18 …”

“…15,16 At the inside of the pores, positively and hydrophobically active ligands can bind the process related impurities like host cell proteins and DNA. 6,17,18 The live recombinant measles vaccine vector technology has multiple advantages: it utilizes the measles virus (MV) vaccine Schwarz strain, a live-attenuated negative-stranded RNA virus, which is one of the safest and most effective human vaccines. A reverse genetic system for the rescue of the negative-stranded MV genome was established.…”