High‐Performance Liquid Chromatography of Peptides and Proteins

Boysen, Reinhard I; Hearn, Milton Thomas William

doi:10.1002/9783527631841.ch5

Cited by 3 publications

(1 citation statement)

References 114 publications

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“…From here on, the maturation of the field was mainly driven by the introduction of analytical and preparative reversed-phase high-performance liquid chromatography [12] and mass spectrometry (MS) techniques such as matrix-assisted linear desorption-ionisation Time-Of-Flight, MALDI TOF MS [13], and liquid chromatography, LC-MS [14], which made it possible for most laboratories to purify and characterize their peptide products.…”

Section: Glumentioning

confidence: 99%

Peptides, Antibodies, Peptide Antibodies and More

Trier

Hansen

Houen

2019

IJMS

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The applications of peptides and antibodies to multiple targets have emerged as powerful tools in research, diagnostics, vaccine development, and therapeutics. Antibodies are unique since they, in theory, can be directed to any desired target, which illustrates their versatile nature and broad spectrum of use as illustrated by numerous applications of peptide antibodies. In recent years, due to the inherent limitations such as size and physical properties of antibodies, it has been attempted to generate new molecular compounds with equally high specificity and affinity, albeit with relatively low success. Based on this, peptides, antibodies, and peptide antibodies have established their importance and remain crucial reagents in molecular biology.

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Section: Glumentioning

confidence: 99%

Peptides, Antibodies, Peptide Antibodies and More

Trier

Hansen

Houen

2019

IJMS

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Salts employed in hydrophobic interaction chromatography can change protein structure – insights from protein‐ligand interaction thermodynamics, circular dichroism spectroscopy and small angle X‐ray scattering

Komaromy

Kulsing

Boysen

et al. 2015

Biotechnology Journal

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Key requirements of protein purification by hydrophobic interaction chromatography (HIC) are preservation of the tertiary/quaternary structure, maintenance of biological function, and separation of the correctly folded protein from its unfolded forms or aggregates. This study examines the relationship between the HIC retention behavior of hen egg white lysozyme (HEWL) in high concentrations of several kosmotropic salts and its conformation, assessed by circular dichroism (CD) spectroscopy. Further, the physicochemical properties of HEWL in the presence of high concentrations of ammonium sulfate, sodium chloride and magnesium chloride were investigated by small angle X-ray scattering (SAXS) at different temperatures. Radii of gyration were extrapolated from Guinier approximations and the indirect transform program GNOM with protein-protein interaction and contrast variation taken into account. A bead model simulation provided information on protein structural changes using ab initio reconstruction with GASBOR. These results correlated to the secondary structure content obtained from CD spectroscopy of HEWL. These changes in SAXS and CD data were consistent with heat capacity ΔCp -values obtained from van't Hoff plot analyses of the retention data. Collectively, these insights enable informed decisions to be made on the choice of chromatographic conditions, leading to improved separation selectivity and opportunities for innovative column-assisted protein refolding methods.

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