Estimation of band broadening in size-exclusion chromatography. I. A method based on analyzing narrow standards with a molar mass-sensitive detector

Yossen, Mariana Matilde; Vega, Jorge Rubén; Meira, G. R.

doi:10.1016/j.chroma.2006.06.062

Cited by 15 publications

(10 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a result, the impact of band broadening in SEC is of particular importance and has been the subject of a wide number of articles. [54,66,70–72] …”

Section: Instrumentation Considerationsmentioning

confidence: 99%

A Review Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and Their Aggregates

Hong

Koza

Bouvier

2012

Journal of Liquid Chromatography & Related Technologies

436

291

View full text Add to dashboard Cite

In recent years, the use and number of biotherapeutics has increased significantly. For these largely protein-based therapies, the quantitation of aggregates is of particular concern given their potential effect on efficacy and immunogenicity. This need has renewed interest in size-exclusion chromatography (SEC). In the following review we will outline the history and background of SEC for the analysis of proteins. We will also discuss the instrumentation for these analyses, including the use of different types of detectors. Method development for protein analysis by SEC will also be outlined, including the effect of mobile phase and column parameters (column length, pore size). We will also review some of the applications of this mode of separation that are of particular importance to protein biopharmaceutical development and highlight some considerations in their implementation.

show abstract

“…As a result, the impact of band broadening in SEC is of particular importance and has been the subject of a wide number of articles. [54,66,70–72] …”

Section: Instrumentation Considerationsmentioning

confidence: 99%

A Review Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and Their Aggregates

Hong

Koza

Bouvier

2012

Journal of Liquid Chromatography & Related Technologies

436

291

View full text Add to dashboard Cite

show abstract

“…One such technique is fluorophore-assisted capillary electrophoresis, − but this is limited by being only useful for low degrees of polymerization (∼100). Another technique is the use of ultra-narrow standards ,,− or even uniform (perfectly monodisperse) standards whereby the broadening function can be determined directly as the signal, using eq 2. However, preparation of ultranarrow or uniform standards is extremely laborious, and none is available commercially.…”

Section: Introductionmentioning

confidence: 99%

Toward a More General Solution to the Band-Broadening Problem in Size Separation of Polymers

et al. 2007

View full text Add to dashboard Cite

The molecular weight distributions (MWDs) and hydrodynamic volume distributions of polymers can reveal considerable mechanistic information on the polymerization process, and have significant effects on physical properties such as viscosity. While the broadening function for a particular SEC setup can be found using ultranarrow standards, these are extremely difficult to obtain. The present paper implements and tests a suggested technique (Aust. J. Chem. 2005, 58, 178) to enable the deconvolution of size distributions using broad standards, synthesized under conditions which are expected to produce a number MWD P(M) which is a single exponential. Broad standards with a wide range of M̄ n were synthesized for both styrene and methyl methacrylate (MMA), using low-conversion free-radical polymerization with appropriate choice of chain transfer agent (CTA) and initiator concentrations; standards with high M̄ nwere synthesized at 25 °C without added initiator. The broadening function was obtained by assuming a flexible functional form (exponential Gaussian hybrid) and least-squares fitting its parameters so that the “theoretical” exponential P(M) curves for each sample, with exponents obtained experimentally, matched the experimental SEC distribution for styrene. The procedure was tested by using the same band-broadening function to deconvolute data for the original polystyrene “standards” and the polyMMA samples, using the Ishige deconvolution method. This method tends to amplify noise, and too tight a tolerance can lead to spurious structure in the deconvoluted distributions. Nevertheless, a tolerance range could be found which led to stable solutions, where the deconvoluted P(M) curves for both were indeed single exponential over the range of molecular weights where data with acceptable accuracy could be obtained. This suggests that this is a generally applicable method to correct for band broadening for a wide range of systems, although improved deconvolution methods are needed to obtain truly converged and stable solutions.

show abstract

“…The two options to circumvent the above described problems are either to correct for chain length dependent band broadening in SEC according to methods described by several authors [39][40][41] or resort to the use of MALDI-ToF MS, possibly in combination with SEC where fractions of the SEC are analyzed with MALDI in order to get the information and the amounts of chains of a particular length by the mass detectors in SEC (for example practiced in the group of Kowolik in Karlsruhe, describes elsewhere in this volume).…”

mentioning

confidence: 99%

Modeling of Emulsion Polymerization, Will It Ever be Possible ? Part‐2: Determination of Basic Kinetic Data Over the Last Ten Years

Herk¹

2009

Macromolecular Symposia

View full text Add to dashboard Cite

Summary: Modeling of Emulsion polymerization processes is based on the knowledge of fundamental kinetic and thermodynamic parameters. The most important kinetic parameters are propagation and termination rate coefficients. Ten years ago a paper has been published with the title ''Modeling of Emulsion Polymerization, will it ever be possible?''. The paper expresses serious doubts whether the different kinetic parameters will ever be available for general industrial recipes containing several monomers. In this paper the developments in the last 10 years regarding pulsed initiation polymerization (PIP) and other techniques regarding the determination of especially the kinetic parameters will be discussed. Amongst the areas where progress can be seen is the understanding of acrylate systems and water soluble monomers. New techniques like MALDI-ToF MS and time resolved electron spin resonance have strongly contributed to the area. Also the puzzling effect of chain length dependent propagation is discussed. Finally an answer will be given to the title question.

show abstract

Estimation of band broadening in size-exclusion chromatography. I. A method based on analyzing narrow standards with a molar mass-sensitive detector

Cited by 15 publications

References 31 publications

A Review Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and Their Aggregates

A Review Size-Exclusion Chromatography for the Analysis of Protein Biotherapeutics and Their Aggregates

Toward a More General Solution to the Band-Broadening Problem in Size Separation of Polymers

Modeling of Emulsion Polymerization, Will It Ever be Possible ? Part‐2: Determination of Basic Kinetic Data Over the Last Ten Years

Contact Info

Product

Resources

About