High throughput process development workflow with advanced decision-support for antibody purification

Stamatis, Christos; Goldrick, Stephen; Gruber, David; Turner, Richard; Titchener‐Hooker, Nigel J.; Farid, Suzanne S.

doi:10.1016/j.chroma.2019.03.005

Cited by 14 publications

(8 citation statements)

References 34 publications

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“…The collected solution was saved as the first elution fraction and repeated this step 3 times to obtain 4 purified fractions. Fractions containing purified antibodies and content were determined by measuring the relative absorbance of each fraction at 280 nm [ 10 , 11 ].…”

Section: Methodsmentioning

confidence: 99%

Purification and Analysis of the CREPT Antibody from Mouse Ascites

Xie

Hao

et al. 2022

Applied Bionics and Biomechanics

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Background and objective. The cell cycle-related and expression-elevated protein in tumor (CREPT) is overexpressed in several human cancers. Establishing a method for purifying CREPT from mouse ascites is vital for further research. This study was aimed at establishing a method for purifying CREPT from mouse ascites. Methods. Cells were cultivated properly to obtain 3E10 CREPT monoclonal antibody cells in the logarithmic growth stage. Monoclonal antibody cells were injected into the abdominal cavity of sensitized mice. The flowing ascites were observed for 7-15 days. The antibody protein was obtained by collection, filtration, dilution, loading, and chromatography. Furthermore, its binding force was detected by SDS-PAGE and Western blot techniques. Results. The antibody protein was successfully obtained with a purity of 1895 μg/mL with high liveness. Conclusion. This study establishes a one-step purification method for obtaining monoclonal antibody with high liveness and purity for CREPT ascites antibody. This method is simple to perform and lays a foundation for the preparation and purification of humanized monoclonal antibodies in the future. In addition, it provides a basis for further research to investigate how CREPT affects the occurrence and development of different tumors.

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Section: Methodsmentioning

confidence: 99%

Purification and Analysis of the CREPT Antibody from Mouse Ascites

Xie

Hao

et al. 2022

Applied Bionics and Biomechanics

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“…An HT method based on a microtiter filter plate [96well with MultiScreen HTS Vacuum Manifold (Merck-Millipore)] was applied to determine the adsorption properties and evaluate the optimal conditions for human serum albumin (HSA) isolation with four MM resins and two IEX resins; the findings were verified by laboratory-scale column chromatography (Chu et al, 2018). In another study, an HT process development workflow integrated with a microscale chromatography, DoE, and multivariate data analysis was studied and provided a rational method for screening resins and process parameters (Stamatis et al, 2019). In a study by Andar et al (2019), a microscale column using IMAC was used for the purification of G-CSF expressed using a cell-free CHO and was compared with a 1 ml IMAC column.…”

Section: High-throughput Technologiesmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

400

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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“…Automation benefits from shortened process development times, limited need of manual intervention, and lower material consumption. During downstream processing, automation is currently utilized for high-throughput (HT) experiments [2,3], liquid and solid dosing steps [4,5] as well as for single unit operations [6,7] and for established processes [8].…”

Section: Introductionmentioning

confidence: 99%

Development of an Automated Adsorbent Selection Strategy for Liquid–Phase Adsorption

Schreiber

Schembecker

2022

Chem Eng & Technol

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A systematic and automatic approach for adsorbent selection for liquid–phase adsorption is proposed. Based on physical properties like polarity, pore size, and specific surface, a screening strategy is developed and automated on a robotic platform. Key performance indicators are applied ensuring economically based decisions. The approach developed is verified by adsorption of caffeine out of an aqueous solution with vanillin, uracil, and α–ionon as impurities. The adsorbent selection strategy leads to the polymeric adsorbent SP207 and a specific surface of 15 m2mL−1 ending in a separation cost indicator of 16 € gCaffeine−1. This work proposes an opportunity for accelerated process design strengthened by the usage of robotic devices.

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High throughput process development workflow with advanced decision-support for antibody purification

Cited by 14 publications

References 34 publications

Purification and Analysis of the CREPT Antibody from Mouse Ascites

Purification and Analysis of the CREPT Antibody from Mouse Ascites

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Development of an Automated Adsorbent Selection Strategy for Liquid–Phase Adsorption

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