Purification activation peptide

Schroder JM, Mrowietz U, Morita E, Christophers E. Purification and partial biochemical characterization of a human monocyte-derived, neutrophil-activating peptide that lacks interleukin 1 activity. J Immunol 1987 139 3474-3483. 

With the advent of HPLC, a new tool was born that revolutionized peptide/protein chemistry as a whole as it allowed not only purification of biologically active peptides and proteins from complex mixtures of tissue or plant extracts/231 but also allowed purification of synthetic unprotected peptide mixtures analytically and preparatively.124-261 This was particularly significant in that purity could be assessed of peptide intermediates made by the classical solution-phase methodology that promoted characterization of all intermediates, as well as the purity of final products made by the solid-phase approach of MerrifieldJ27 ... 

Clark-Lewis, I., Moser, B., Walz, A., Baggiolini, M Scott, G. J., and Aebersold, R. (1991) Chemical synthesis, purification, and characterization of two inflammatory proteins, neutrophil activating peptide I (interleukin-8) and neutrophil activating peptide 2. Biochemistry 30, 3128. 

Schroder, J. M. (1997) Isolation and purification of neutrophil-activating peptide-4 a chemokine missing two cysteines. Methods Enzymol. 287, 216. 

S. Masure, P. Proost, J. van Damme, and G. Opdenakker, Purification and Identification of9l-lcDa Neutrophil Gelatinase Release by the Activating Peptide Interleukin-8, European Journal of Biochemistry, 198 (1991) 391 -398. 

Affinity purification of peptides by immobilized enzymes relies on the highly specific interaction of an ionic, hydrophobic, aromatic, or stoically active binding site on the desired peptide to the immobilized enzyme. The unbound contaminants are washed through and the desired peptide is released by buffer elution. This technique is beneficial in the purification of many peptides and can also be used for affinity chromatog-... 

Results of the separation of the activation peptides, obtained as described above, are shown in Figure 3. Amino acid analysis of the peaks showed that the pepstatin was recovered in peak 1 and that peak 4 contained mainly peptide 1-16 (Table I). In addition to the amino acids listed, there were small amounts of histidine, glycine, and alanine. From the quantities of these amino acids and the known sequence of the total activation fragments (5,21,22), it was ascertained that the product was about 90% pure, the slightly high values for several amino acids being caused by contaimination with at least three other peptides. No further purification was considered considered necessary. 

---