UNIGE document Scientific Article
previous document  unige:101435  next document
add to browser collection
Title

Influence of connection tubing in modern size exclusionchromatography and its impact on the characterization of mAbs

Authors
Published in Journal of Pharmaceutical and Biomedical Analysis. 2018, vol. 149, p. 22-32
Abstract The goal of the study was to evaluate the impact of connection tubing in modern size exclusion chro-matography (SEC), since it may strongly impact the apparent column efficiency, as the compounds arenot retained in SEC. For this purpose, a reference SEC column of 150 × 4.6 mm, 1.8 m was considered,and various proteins were tested as model compounds. Different tube geometries (lengths and internaldiameters) and materials (stainless steel and PEEK) were evaluated in a systematic way. Large proteinsalways showed larger tube dispersion vs. small molecules, especially when the residence time in thetube was long (at low flow rate). This confirms the need to drastically reduce the tube volume (using the shortest and narrowest connector tubing) to attain the full benefits of UHPSEC columns. In addition,PEEK tubing were found to be more appropriate than stainless steel tubing, since adsorption of proteinswas less pronounced, and higher plate count can be obtained. Finally, after a careful system optimization,up to 40% increase of apparent column efficiency can be achieved compared to a regular UHPLC system,when using a 150 × 4.6 mm UHPSEC columns packed with sub–3 m particles.
Keywords Size exclusion chromatographyConnection tubingExtra-column band broadeningProteinsMonoclonal antibodySub–3 m SEC column
Identifiers
PMID: 29100027
Full text
Article (Published version) (3.3 MB) - document accessible for UNIGE members only Limited access to UNIGE
Structures
Research group Sciences analytiques
Citation
(ISO format)
FEKETE, Szabolcs, GUILLARME, Davy. Influence of connection tubing in modern size exclusionchromatography and its impact on the characterization of mAbs. In: Journal of Pharmaceutical and Biomedical Analysis, 2018, vol. 149, p. 22-32. https://archive-ouverte.unige.ch/unige:101435

61 hits

0 download

Update

Deposited on : 2018-01-16

Export document
Format :
Citation style :