9/20/2023 0 Comments Capto blue![]() ![]() Īpart from cost savings, the re‐use of chromatography resins can be a necessity in situations wherein supply limitations are in place. ![]() The number of cycles can vary from small to large in batch and continuous processes and column re‐use in such processes requires the completion of studies validating the lifespan of chromatography media. This typically includes the deployment of multiple CIP agents, such as high conductivity and caustic solutions, aiming to remove tightly bound residuals from the resin that would otherwise lead to its fouling and to a potentially significant reduction of the resin's performance in purifying a target product over multiple cycles. The development of CIP strategies for biopharmaceuticals, such as monoclonal antibodies, has been reported for affinity, ion exchange, and hydrophobic interaction‐ion exchange (e.g., ) resin modalities. 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.Ībbreviations Au absorbance units BSA bovine serum albumin CCCH clarified cell culture harvest ChromP chromatography flowthrough product pool CIP Cceaning in place CIP1 cleaning in place step 1 CIP2 cleaning in place step 2 CIP3 cleaning in place step 3 CV column volume DBC 10% dynamic binding capacity at 10% breakthrough DF diafiltration FBS fetal bovine serum FT flowthrough (effluent collected during column loading) G V vesicular stomatitis virus glycoprotein hcDNA host cell DNA HCP host cell proteins HT high throughput L vesicular stomatitis virus large polymerase LMH liters per square meter hour LOQ limit of quantitation LVV live virus vaccine M vesicular stomatitis virus matrix protein N vesicular stomatitis virus nucleoprotein protein P vesicular stomatitis virus phosphoprotein PAGE polyacrylamide gel electrophoresis PBS phosphate buffered saline PCIP post cleaning in place Pen Strep Penicillin‐Streptomycin qPCR quantitative polymerase chain reaction RC RoboColumn S SARS‐CoV‐2 spike glycoprotein SDS sodium dodecyl sulfate TEM transmission electron microscopy UF ultrafiltration UFCR ultrafiltration concentrated retentate UFDR ultrafiltration diafiltrated retentate UFP ultrafiltration product V 10% 10% breakthrough volume V o column void volume VSV vesicular stomatitis virus ρ Spearman's rank correlation coefficientĬost of goods for bioprocess is typically dominated by downstream processing and hence the implementation of cleaning in place (CIP) techniques for chromatography resins is important for mitigating the costs associated with the use of this unit operation. 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. Here, its impact was assessed across the entire purification process which also included an ultrafiltration/diafiltration step. The success of the CIP strategy was ultimately verified at the laboratory scale. ![]() 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. 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. 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. ![]()
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