High performance liquid chromatography of proteins on a diol-bonded silica gel stationary phase

Schmidt, Donald E.; Giese, Roger W.; Conron, D.W.; Karger, Barry L.

doi:10.1021/ac50051a040

Cited by 126 publications

(22 citation statements)

References 27 publications

(28 reference statements)

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“…an extensive characterization of the functional properties of a TSK 3000SW column*. These studies confirm the large effect that the residual negative charge of the matrix can have on protein migration at low ionic strength (7,9). We have extended these observations by demonstrating that with a high enough ionic strength, protein migration is not only independent of column charge, but is also consistent with the elution characteristics expected based on the protein's molecular weight and the hydrodynamic Stokes radius.…”

mentioning

confidence: 65%

“…However, their results also showed that salts such as Na2S04, which have more powerful salting-out properties, can produce retardation of proteins due to a "hydrophobic-type interaction." Lysozyme has been suggested to interact with this column and other HPSEC packings via a hydrophobic mechanism (7). This mechanism probably explains some of the retardation of lysozyme on the TSK column (F, = 1.00); however, lysozyme has a very high PI (2 11.0) and may be retarded electrostatically.…”

Section: Discussionmentioning

confidence: 94%

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Use of high‐performance size‐exclusion chromatography to measure protein molecular weight and hydrodynamic radius

Tarvers

Church

1985

International Journal of Peptide and Protein Research

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We have conducted a study of the TSK 3000 SW high‐performance size‐exclusion column to define under what conditions proteins would migrate most consistently with their known hydrodynamic properties. Our findings include the following: 1) the residual negative charge of the column does cause charge‐exclusion or charge‐retention effects at low ionic strengths; with elution in deionized water several anionic proteins elute approximately in the void volume; 2) at μ > 0.5, protein migration is not only independent of ionic strength, but consistent with protein molecular weight and hydrodynamic volume; 3) small hydrophobic peptides are retarded by the column; and 4) very asymmetric proteins and other hydrodynamic particles are likely to be retarded by an “end‐on insertion” mechanism.

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mentioning

confidence: 65%

Section: Discussionmentioning

confidence: 94%

Use of high‐performance size‐exclusion chromatography to measure protein molecular weight and hydrodynamic radius

Tarvers

Church

1985

International Journal of Peptide and Protein Research

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“…(15) and the protein is assumed to be excluded from the pores (80 A.) as previously demonstrated by Schmidt et al (16) on LiChrosorb Diol 100 A. HSA would thus be mainly adsorbed on the external surface of the RP support.…”

Section: Adsorption Of Usa In Aqueous Buffermentioning

confidence: 94%

Study of the Adsorption of Human Serum Albumin on Reversed-Phase Supports.

Place¹,

Sébille²,

Vidal-Madjar³

1992

Journal of Dispersion Science and Technology

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“…Conditions for column chromatography were as described in the legend to Fig. 1 governed by (1) hydrophobic interaction between the solute and the hydrocarbon ligand of the stationary phase (Horvath et al, 1976) and (2) ionic interaction between the charged solute with the negatively charged silica surface of the stationary phase (Schmidt et al, 1980 The gradient system described in Fig. 1 Sequence analysis ofpolypeptide The method has been applied to sequence determination of polypeptides with both known and unknown structures.…”

Section: Preparation Of By-products Of Dabth-ser and Dabth-thrmentioning

confidence: 99%

N-terminal sequence analysis of polypeptide at the picomole level

Chang¹

1981

Biochemical Journal

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This paper describes a manual method for N-terminal sequence analysis of polypeptides at subnanomole sensitivity. The polypeptide is degraded stepwise by using the dimethylaminoazobenzene isothiocyanate/phenyl isothiocyanate double-coupling method, and the released dimethylaminoazobenzenethiohydantoins of amino acids were identified by reversed-phase high-pressure liquid chromatography. The dimethylaminoazobenzenethiohydantoins are coloured compounds and can be detected in the visible region with the sensitivity limit of 1 pmol (signal-to-baseline noise ratio 5). A high-pressure liquid-chromatographic method was developed for complete analysis of all amino acid dimethylaminoazobenzenethiohydantoin derivatives, including the byproducts of serine and threonine. Thus, without use of an automatic sequenator or radioactive materials, it is possible to determine the complete sequence of peptides and N-terminal sequence of proteins with less than 1 nmol of material.

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High performance liquid chromatography of proteins on a diol-bonded silica gel stationary phase

Cited by 126 publications

References 27 publications

Use of high‐performance size‐exclusion chromatography to measure protein molecular weight and hydrodynamic radius

Use of high‐performance size‐exclusion chromatography to measure protein molecular weight and hydrodynamic radius

Study of the Adsorption of Human Serum Albumin on Reversed-Phase Supports.

N-terminal sequence analysis of polypeptide at the picomole level

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