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

Journal of Pharmaceutical and Biomedical Analysis
Szabolcs Fekete, Davy Guillarme

Abstract

The goal of the study was to evaluate the impact of connection tubing in modern size exclusion chromatography (SEC), since it may strongly impact the apparent column efficiency, as the compounds are not retained in SEC. For this purpose, a reference SEC column of 150×4.6mm, 1.8μm was considered, and various proteins were tested as model compounds. Different tube geometries (lengths and internal diameters) and materials (stainless steel and PEEK) were evaluated in a systematic way. Large proteins always showed larger tube dispersion vs. small molecules, especially when the residence time in the tube 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 proteins was 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 150×4.6mm UHPSEC columns packed with sub-3μm particles.

References

Jul 14, 2004·Journal of Chromatography. a·G GrznárováT Görner
Jul 22, 2004·Analytical Biochemistry·J JenkinsS Sundaram
Feb 5, 2013·Journal of Liquid Chromatography & Related Technologies·Paula HongEdouard S P Bouvier
Apr 11, 2013·Journal of Pharmaceutical and Biomedical Analysis·Szabolcs FeketeDavy Guillarme
May 13, 2014·Journal of Pharmaceutical and Biomedical Analysis·Szabolcs FeketeDavy Guillarme
Jun 14, 2014·Journal of Chromatography. a·Fabrice Gritti, Georges Guiochon
Nov 26, 2015·Analytical Chemistry·Jelle De VosSebastiaan Eeltink
May 9, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Balázs BobályJenő Fekete
Dec 20, 2016·Journal of Pharmaceutical and Biomedical Analysis·Alexandre GoyonSzabolcs Fekete

❮ Previous
Next ❯

Citations

Feb 29, 2020·Annual Review of Analytical Chemistry·Charles R Bupp, Mary J Wirth
Jun 25, 2018·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Alexandre GoyonDavy Guillarme

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.