Slalom chromatography (SC) is an alternative chromatographic procedure for the separation of relatively large double-stranded DNA molecules and is based on a new principle. The retardation of the DNA fragments from the cleavage of the Lambda DNA by the KpnI restriction enzyme was studied using an acetonitrile-phosphate buffer as a mobile phase with various concentrations of viscosity modifier (i.e. glycerol) and a C1 column as a stationary phase. The DNA molecule retention was accurately described over the glycerol concentration range using a model previously established. It was shown that the eluent viscosity increase enhanced the slalom chromatographic capacity to separate the DNA fragments. A connection between SC and 'hydrodynamic chromatography' processes was predicted to link the two processes in a global separation mechanism based on a non-equilibrium principle.
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Ultra-high pressure slalom chromatography: Application to the characterization of large DNA and RNA samples relevant in cell and gene therapy.
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Waters Corporation, Core Research/Fundamental Milford, MA, 01757, USA. Electronic address:
Slalom chromatography (SC), initially co-discovered by Boyes and Kasai in the late 1980s, has recently re-emerged as a breakthrough technique to rapidly analyze DNA samples. With the development of cutting-edge ultra-high pressure liquid chromatography (UHPLC) systems and columns, SC now offers enhanced resolution and sensitivity for analyzing large DNA samples. By revisiting the fundamentals of the SC retention mechanism (non-equilibrium separation mode) and considering the physicochemical properties of DNA biopolymers (contour length, extension under shear flow, relaxation time), we provide analytical chemists with a general strategy and framework for selecting the most relevant applications in the expanding field of cell and gene therapy.
View Article and Find Full Text PDFTheoretical predictions to facilitate the method development in slalom chromatography for the separation of large DNA molecules.
J Chromatogr A
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Waters Corporation, Core Research/Fundamental, Milford, MA, 01757, USA. Electronic address:
Slalom chromatography (SC) re-emerged in 2024 due to the availability of low adsorption ultra-high pressure liquid chromatography (UHPLC) packed columns/instruments and large modalities being investigated in the context of cell and gene therapies. The physico-chemical principles of SC retention combined with hydrodynamic chromatography (HDC) exclusion have been recently reported. In SC, DNA macromolecules are retarded because: (1) they can be stretched to lengths comparable to the particle diameter, and (2) their elastic relaxation time is long enough to maintain them in non-equilibrium extended conformations under regular UHPLC shear flow conditions.
View Article and Find Full Text PDFRetention mechanism in combined hydrodynamic and slalom chromatography for analyzing large nucleic acid biopolymers relevant to cell and gene therapies.
J Chromatogr A
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Waters Corporation, Instrument/Core Research/Fundamental, Milford, MA, 01757, USA.
Slalom chromatography (SC) was discovered in 1988 for analyzing double-stranded (ds) DNA. However, its progress was impeded by practical issues such as low-purity particles, sample loss, and lack of a clear retention mechanism. With the rise of cell and gene therapies and the availability today of bio-inert ultra-high-pressure liquid chromatography (UHPLC) columns and systems, SC has regained interest.
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Carbohydr Polym
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Department of Chemistry and Polymer Science, University of Stellenbosch, Private Bag X1, 7602 Matieland, South Africa.
Elution behavior of chitosan was studied using narrow-bore size exclusion chromatography (SEC) columns packed with 8 μm porous particles at different eluent flow rates. Delayed elution of the largest chitosan species from the columns was observed at high flow rates, which was attributable to strong chain deformation rather than chain degradation. In particular, a chromatographic mode transition from SEC to slalom chromatography (SC) was observed for high molar mass chitosan samples with abnormal elution behaviors, which originates from the coil-stretch transition of chitosan chains in elongational flow through the columns.
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State Key Laboratory of Polymer Physics and Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China. Electronic address:
Here we present SEC-MALLS study for chitosan samples with weight average molar mass M between 33 and 427 kg/mol on columns packed with 8 μm porous particles. A low injection concentration on the order of 0.1-0.
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