Development of affinity chromatography using a bioactive peptide as a ligand.
By repeatedly introducing hydrophilic polyethylene glycol (PEG) spacer (2) onto affinity resin bearing a bioactive peptide (1/2 secretory leukocyte protease inhibitor, 1/2SLPI) as a ligand, the adsorption of nonspecific binding proteins was effectively reduced and the purification efficacy of elastase, which is one of the target molecules for 1/2SLPI, from a protein mixture was improved. Moreover, using this resin, we also successfully detected L-plastin, as an endogenous target molecule for SLPI, from HL-60 cell lysate.
Furuya
Minoru
M
Pharmaceutical Discovery Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Ltd, 4-3-2 Asahigaoka, Hino, Tokyo 191-8512, Japan. mi.furuya@teijin.co.jp
Tsushima
Yu
Y
Tani
Shinobu
S
Kamimura
Takashi
T
D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods; D018922:HL-60 Cells; D006801:Humans; D008024:Ligands; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010446:Peptide Fragments / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry / Q000302:isolation & purification; D011092:Polyethylene Glycols / Q000737:chemistry; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000737:chemistry*; D053481:Secretory Leukocyte Peptidase Inhibitor; D015842:Serine Proteinase Inhibitors / Q000737:chemistry; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000737:chemistry; D013499:Surface Properties
D008024:Ligands; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; C506814:SLPI protein, human; D053481:Secretory Leukocyte Peptidase Inhibitor; D015842:Serine Proteinase Inhibitors; C026742:plastin; D011092:Polyethylene Glycols