Separation of proteins by reversed-phase high-performance liquid chromatography

Qualitative and quantitative characterization of the arsenic‐binding behaviour of sulfur‐containing peptides and proteins by the coupling of reversed phase liquid chromatography to electrospray ionization mass spectrometry

“…Different conformers that arise from changing chemical environment during gradient elution occur in the form of multiple peaks. [22,23] In agreement with the former RPLC-UV analyses in which the water-acetonitrile gradient elution was supported by an ion pairing reagent, [15] the phenylarsenic-substituted peptide forms elute later than the unsubstituted forms owing to a larger molecular size.…”

Qualitative and quantitative characterization of the arsenic‐binding behaviour of sulfur‐containing peptides and proteins by the coupling of reversed phase liquid chromatography to electrospray ionization mass spectrometry

“…Different conformers that arise from changing chemical environment during gradient elution occur in the form of multiple peaks. [22,23] In agreement with the former RPLC-UV analyses in which the water-acetonitrile gradient elution was supported by an ion pairing reagent, [15] the phenylarsenic-substituted peptide forms elute later than the unsubstituted forms owing to a larger molecular size.…”

Superficially porous silica particles with wide pores for biomacromolecular separations

“…Native and non-native forms can exist simultaneously, so that in the reversed-phase separation process the usual approach is to ensure that the protein is in a stable chromatographic configuration, usually in a non-native state. A mobile phase with trifluoroacetic acid or containing urea or some other agent often is used in an attempt to produce stable configurations for separating proteins [24]. …”

A Targeted LC‐MS Strategy for Low‐Abundant HLA Class‐I‐Presented Peptide Detection Identifies Novel Human Papillomavirus T‐Cell Epitopes

“…The most common detergents are CHAPS and IGEPAL CA‐630. Detergents have to be removed before downstream LC‐MS analysis as they significantly influence peptide separation on the LC column53, 54 or interfere with ionization of analytes 55, 56. Therefore, thorough and repeated washing of IP samples is necessary.…”

Qualitative and quantitative characterization of the arsenic‐binding behaviour of sulfur‐containing peptides and proteins by the coupling of reversed phase liquid chromatography to electrospray ionization mass spectrometry

“…Different conformers that arise from changing chemical environment during gradient elution occur in the form of multiple peaks. [22,23] In agreement with the former RPLC-UV analyses in which the water-acetonitrile gradient elution was supported by an ion pairing reagent, [15] the phenylarsenic-substituted peptide forms elute later than the unsubstituted forms owing to a larger molecular size.…”

Superficially porous silica particles with wide pores for biomacromolecular separations

“…Native and non-native forms can exist simultaneously, so that in the reversed-phase separation process the usual approach is to ensure that the protein is in a stable chromatographic configuration, usually in a non-native state. A mobile phase with trifluoroacetic acid or containing urea or some other agent often is used in an attempt to produce stable configurations for separating proteins [24]. …”

A Targeted LC‐MS Strategy for Low‐Abundant HLA Class‐I‐Presented Peptide Detection Identifies Novel Human Papillomavirus T‐Cell Epitopes

“…The most common detergents are CHAPS and IGEPAL CA‐630. Detergents have to be removed before downstream LC‐MS analysis as they significantly influence peptide separation on the LC column53, 54 or interfere with ionization of analytes 55, 56. Therefore, thorough and repeated washing of IP samples is necessary.…”

Qualitative and quantitative characterization of the arsenic‐binding behaviour of sulfur‐containing peptides and proteins by the coupling of reversed phase liquid chromatography to electrospray ionization mass spectrometry

“…Different conformers that arise from changing chemical environment during gradient elution occur in the form of multiple peaks. [22,23] In agreement with the former RPLC-UV analyses in which the water-acetonitrile gradient elution was supported by an ion pairing reagent, [15] the phenylarsenic-substituted peptide forms elute later than the unsubstituted forms owing to a larger molecular size.…”

Superficially porous silica particles with wide pores for biomacromolecular separations

“…Native and non-native forms can exist simultaneously, so that in the reversed-phase separation process the usual approach is to ensure that the protein is in a stable chromatographic configuration, usually in a non-native state. A mobile phase with trifluoroacetic acid or containing urea or some other agent often is used in an attempt to produce stable configurations for separating proteins [24]. …”

A Targeted LC‐MS Strategy for Low‐Abundant HLA Class‐I‐Presented Peptide Detection Identifies Novel Human Papillomavirus T‐Cell Epitopes

“…The most common detergents are CHAPS and IGEPAL CA‐630. Detergents have to be removed before downstream LC‐MS analysis as they significantly influence peptide separation on the LC column53, 54 or interfere with ionization of analytes 55, 56. Therefore, thorough and repeated washing of IP samples is necessary.…”