Dissociation of Antibody–Contaminant Complexes With Hydroxyapatite

Gagnon, Pete

doi:10.12665/j92.gagnon

Cited by 8 publications

(2 citation statements)

References 13 publications

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“…While some of the DNA is bound to hitchhiking H1, the rest appears to bind to conserved, relatively alkaline regions found on most antibodies. Such non-specific complexation of DNA directly to the antibody structure, independent of CDRs, has been observed on IgGs [15] as well as on IgMs [16].…”

Section: The Hitchhiker Threat To Researchmentioning

confidence: 90%

The effects of hitchhiker antigens co-eluting with affinity-purified research antibodies

Mechetner

Sood

Nguyen

et al. 2011

Journal of Chromatography B

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Section: The Hitchhiker Threat To Researchmentioning

confidence: 90%

The effects of hitchhiker antigens co-eluting with affinity-purified research antibodies

Mechetner

Sood

Nguyen

et al. 2011

Journal of Chromatography B

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“…As a third criterion, the qualitative nucleic acid factor NF was determined by the ratio of the 254 nm signal and 280 nm signal. In contrast to measurements at 260 nm, DNA and RNA show almost equivalent absorption at these wavelengths, whereas proteins exhibit a lower absorption at 254 nm and, hence, can be distinguished better from nucleic acids . The factor is defined to be the ratio between the integrated 254 nm signal A 254 and 280 nm signal A 280 (Eq.…”

Section: Methodsmentioning

confidence: 99%

Systematic purification of salt‐intolerant proteins by ion‐exchange chromatography: The example of human α‐galactosidase A

Baumann

Osberghaus

Hubbuch

2015

Engineering in Life Sciences

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Chromatography is an essential tool for purifying biopharmaceutical products. Many processes are still developed based on traditional routines and empirical procedures. Product losses are mostly due to insufficient optimization of purification setups and product sensitivity to process conditions. In order to eliminate these shortcomings, a systematic strategy for the setup of ion‐exchange chromatography is presented, which considers both product stability as well as operational conditions. The stages—a hybrid approach combining high‐throughput screening and analytical small‐scale chromatography—are as follows: (1) pH stability (short‐term); (2) pH stability (long‐term), followed by a screening of additives to enhance protein stability, if required; (3) analytical pH gradient chromatography for evaluation of the operational pH window; and (4) salt stability (long‐term) in the operational pH window determined. The efficiency and straightforwardness of the strategy were shown in a case study on capturing the human α‐galactosidase A enzyme. Following the above procedure, the enzyme was found to be salt‐unstable; a purification factor of 13.2, a concentration factor of 4, and an overall yield of 84.3% were achieved. The applied strategy allowed for a quick establishment of a dedicated capture step at low salt concentrations under stable conditions by well‐chosen prior screening experiments.

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Opportunities and Limitations of Disposable Technologies in Biopharmaceutical Processes

Brod

Vester

Kauling

2012

Chemie Ingenieur Technik

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Während der Einsatz einzelner, vorwiegend einfacher Wegwerfkomponenten in Labor, Pilotanlage und Produktionsumfeld längst gängig ist, werden jetzt ganze Pilot‐ oder Produktionsanlagen weitgehend auf Basis von Disposable‐Komponenten konzipiert. Hiermit verbunden sind große Erwartungen bzgl. Zeit‐ und Kosteneinsparungen. Es wird ein Überblick über den heutigen Stand der Technik für das Upstream Processing und die Aufreinigung gegeben. Vor‐ und Nachteile des Einsatzes von Disposable‐Technologie werden für ausgewählte Verfahrensschritte aufgezeigt. Am Beispiel einer Conceptual‐Design‐Studie für eine Anlage zur Herstellung von monoklonalen Antikörpern wird das Potenzial der Disposable‐Technologie diskutiert. Abschließend werden künftige Chancen und Herausforderungen der Disposable‐Technologien kritisch beleuchtet.

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Dissociation of Antibody–Contaminant Complexes With Hydroxyapatite

Cited by 8 publications

References 13 publications

The effects of hitchhiker antigens co-eluting with affinity-purified research antibodies

The effects of hitchhiker antigens co-eluting with affinity-purified research antibodies

Systematic purification of salt‐intolerant proteins by ion‐exchange chromatography: The example of human α‐galactosidase A

Opportunities and Limitations of Disposable Technologies in Biopharmaceutical Processes

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