Advances on Size Exclusion Chromatography and Applications on the Analysis of Protein Biopharmaceuticals and Protein Aggregates: A Mini Review

Brusotti, Gloria; Calleri, Enrica; Colombo, Raffaella; Massolini, Gabriella; Rinaldi, Francesca; Temporini, Caterina

doi:10.1007/s10337-017-3380-5

Cited by 58 publications

(36 citation statements)

References 78 publications

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“…Size exclusion chromatography (SEC) or gel filtration chromatography separates protein molecules based on their molecular weight. SEC is used for the purification of various proteins such as scFv and insulinlike growth factor receptor (Levin et al, 2015) as well as for aggregate removal and desalting (Brusotti et al, 2018;Burgess, 2018).…”

Section: Chromatography Processesmentioning

confidence: 99%

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

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

336

217

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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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Section: Chromatography Processesmentioning

confidence: 99%

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

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

336

217

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“…Das Vorhandensein dieser hoch-affinen Proteine unterhalb einer Schicht aus lose gebundenen Proteinen konnte durch Kombination von Zentrifugation mit anschließender Tr ennung mittels AF4 bestätigt werden. [51] Diese auftretende Scherbeanspruchung verändert hçchstwahrscheinlich die Zusammensetzung der Proteinkorona, indem bestimmte,locker gebundene Proteine oder Proteinschichten entfernt werden. [45] Eine andere,z ur AF4 verwandte,C hromatographie-ähnliche Technik -d ie hydrodynamische Chromatographie (HDC) -t rennt die Probe ebenfalls nach dem hydrodynamischen Volumen.…”

Section: Isolierung Der Proteinkorona Basierend Auf Chromatographischunclassified

Möglichkeiten und Limitierungen verschiedener Trenntechniken zur Analyse der Proteinkorona

2019

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Die gebildete Proteinkorona ist für das Verhalten von Nanoträgern im biologischen System mitverantwortlich. Die Aufklärung der Bildung und des Einflusses der Proteinkorona auf die biologische Antwort ist daher entscheidend. Ergebnisse der Proteinkorona‐Charakterisierung können jedoch nicht ohne Weiteres untereinander verglichen werden, da es zahlreiche verschiedene Verfahren zur Probenvorbereitung gibt, die jeweils für unterschiedliche Materialien und Fragestellungen geeignet sind. Abhängig von der verwendeten Methode wird der Nanoträger‐Protein‐Komplex auf bestimmte Weise verändert, sodass der individuelle Einfluss der Probenvorbereitung auf die Proteinkorona berücksichtigt werden muss. Ziel dieses Kurzaufsatzes ist es daher, einen Überblick über Probevorbereitungsverfahren für die Analyse der Proteinkorona zu geben und ihren Einfluss auf das Ergebnis zu beurteilen, besonders im Hinblick auf die eingeführten Begriffe “weiche” und “harte” Proteinkorona. Im Fokus liegen dabei vor allem die am häufigsten verwendeten Techniken wie Zentrifugation, magnetische sowie chromatographische Trennungen.

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“…different disulfide bonds)Expensive equipmentInvolatile sample buffers are not compatible with ESI–IMS–MSFuruki et al (2017)Young et al (2016)Extrinsic fluorescenceTertiary and quaternary structureCa. 0.015 g/l for 50 kDa proteinSensitiveSuitable for high-throughput screeningDye might interfere with protein aggregationHawe et al (2008)Younan and Viles (2015)Fourier transform infrared spectroscopy (ATR-FTIR)Secondary structure and dynamics> 0.01 g/lTolerant to salt and sample turbidityCan be used for all proteinsHigh wavelength precisionNo time-related dataPathak et al (2016)Walther et al (2014)Baldassarre and Barth (2014)Nuclear magnetic resonance spectroscopy (NMR)Tertiary and quaternary structure> 25 g/l for 50 kDa proteinReal-time application possibleNon-destructiveStructure can be analyzed under native conditionsLarge number of samples necessaryLimited to small proteins (≤ 40 kDa) or protein fragmentsLanucara et al (2014)Kelly et al (2005)Reversed phase high performance liquid chromatography (RP-HPLC)Monitor unfolded proteins and primary structure> 0.3 g/lRapidHigh resolutionRobustHigh temperature during analysis may lead to aggregate formationMere chemical information because proteins are denaturedSturaro et al (2016)Herman et al (2002)Size exclusion high performance liquid chromatography (SE-HPLC)Quantitative protein analysis0.012 g/lNon-destructiveSensitiveNon-denaturating elution conditions possibleLimited dynamic rangeInaccuracy due to alteration of size distributionCodevilla et al (2004)Brusotti et al (2017)Raman spectroscopySecondary structureProtein quantification (SERS)> 1 g/l> 0.08 g/l for SERSSensitive and structural selectiveExpensive equipment (CW-UV laser) and complicated instrumentationBalakrishnan et al (2008)Eryilmaz et...…”

Section: Monitoring Toolsmentioning

confidence: 99%

Wanted: more monitoring and control during inclusion body processing

Humer

Spadiut

2018

World J Microbiol Biotechnol

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Inclusion bodies (IBs) are insoluble aggregates of misfolded protein in Escherichia coli. Against the outdated belief that the production of IBs should be avoided during recombinant protein production, quite a number of recombinant products are currently produced as IBs, which are then processed to give correctly folded and soluble product. However, this processing is quite cumbersome comprising IB wash, IB solubilization and refolding. To date, IB processing often happens rather uncontrolled and relies on empiricism rather than sound process understanding. In this mini review we describe current efforts to introduce more monitoring and control in IB processes, focusing on the refolding step, and thus generate process understanding and knowledge.

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Advances on Size Exclusion Chromatography and Applications on the Analysis of Protein Biopharmaceuticals and Protein Aggregates: A Mini Review

Cited by 58 publications

References 78 publications

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

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

Möglichkeiten und Limitierungen verschiedener Trenntechniken zur Analyse der Proteinkorona

Wanted: more monitoring and control during inclusion body processing

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