Use of Trialkyl Ammonium Phosphate (TAAP) Buffers in Reverse Phase HPLC for High Resolution and High Recovery of Peptides and Proteins

Abstract A new buffer system compatible with reverse phase high pressure liquid chromatography (RP -HPLC) column packings and giving high recoveries and high resolution for peptides and proteins is described. The TEAP buffer (triethylammonium phosphate), one of several TAAP buffers (trialkylammonium phosphate) compatible with RP-HPLC, gives high resolution and high recovery for closely related peptides or proteins as well as day to day reproducibility. It is UV transparent to less than 200 nm, extremely versatile, and could probably be used for several other classes of compounds (phosphatidyl cholines and sphingomyelin, for example). Furthermore, this system is compatible with in vitro and in vivo biological systems after removal of the organic solvent. We have applied our system to the purification of radioactive peptides, i.e. the separation and total recovery of tritiated luteinizing hormone releasing factor (LRF) from closely related radioactive impurities. Peptides and proteins as different as LRF, s...

[1]  J. Rivier,et al.  NEUROTENSIN ANALOGS. STRUCTURE-ACTIVITY RELATIONSHIPS , 1978 .

[2]  C. Horváth,et al.  Separation of amino acids and peptides on non-polar stationary phases by high-performance liquid chromatography. , 1977, Journal of chromatography.

[3]  J. Rivier,et al.  Neurotensin analogues. Structure--activity relationships. , 1977, Journal of medicinal chemistry.

[4]  G. Cox.  Practical Aspects of Bonded Phase Chromatography , 1977 .

[5]  J. Knox.  Practical Aspects of LC Theory , 1977 .

[6]  C. Horváth,et al.  Liquid Chromatography with Hydrocarbonaceous Bonded Phases; Theory and Practice of Reversed Phase Chromatography , 1977 .

[7]  T. Greibrokk,et al.  High-pressure liquid chromatography of peptides. , 1977, Journal of chromatography.

[8]  R. W. Frei,et al.  The separation of nonapeptides by reversed-phase high-performance liquid chromatography. , 1977, Journal of chromatography.

[9]  W. Hancock,et al.  High pressure liquid chromatography in the analysis of underivatised peptides using a sensitive and rapid procedure , 1976, FEBS letters.

[10]  N. Ling,et al.  Isolation, primary structure, and synthesis of α-endorphin and γ-endorphin, two peptides of hypothalamic-hypophysial origin with morphinomimetic activity , 1976 .

[11]  N. Ling,et al.  Anatomic and phylogenetic distribution of somatostatin. , 1976, Metabolism: clinical and experimental.

[12]  C. L. Zimmerman,et al.  Advances in the analysis of amino acid phenylthiohydantoins by high performance liquid chromatography. , 1976, Analytical biochemistry.

[13]  W. Vale,et al.  Myelin basic protein: a substance that releases immunoreactive growth hormone in vitro. , 1976, Biochemical and biophysical research communications.

[14]  I. Shechter.  Separation of proteins by high-speed pressure liquid chromatography. , 1974, Analytical biochemistry.

[15]  J. Miller,et al.  Photometric analysis of proteins and peptides at 191-194 millimicrons. , 1970, Analytical biochemistry.

[16]  P. R. O'Bar,et al.  Absorption of Proteins and Peptides in the Far Ultraviolet , 1970, Science.

[17]  Gerald L. Hawk,et al.  Biological Biomedical Applications of Liquid Chromatography , 1979 .

[18]  K. Tsuji,et al.  Improved high-performance liquid chromatographic method for polypeptide antibiotics and its application to study the effects of treatments to reduce microbial levels in bacitracin powder. , 1975, Journal of chromatography.

[19]  L. Pickart,et al.  Purification of growth-promoting peptides and proteins, and of histones, by high pressure silica gel chromatography. , 1975, Preparative biochemistry.