Ligand profiling and identification technology for searching bioactive ligands.
We introduce a new methodology named ligand profiling and identification for effective discovery of bioactive ligands such as peptide hormones. This technology was developed from a new concept of parallel column chromatography and active fraction profiling by nano-LC MS. Traditional methods use sequential column chromatography, and thus are inevitably limited by the low abundance of the peptide of interest and by a low yield due to the many column steps. Using this new technology, insulin was successfully identified and diarginylinsulin, a minor intermediate form of insulin, was unexpectedly also identified simultaneously from 100 mg of porcine pancreatic tissue. This integrative technology could be used to search for various low-abundance peptides (or bioactive molecules) rapidly and simultaneously, by applying this to the later stages of traditional sequential purification.
Baek
Moon-Chang
MC
Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea.
Kim
Sun-Jin
SJ
Yea
Kyungmoo
K
Kim
Youndong
Y
Lee
Byung-Dae
BD
Kim
Jaeyoon
J
Lee
Hae-Jeong
HJ
Kang
Mee-Hee
MH
Choi
Sun-Kyu
SK
Kim
Jong-In
JI
Lee
Taehoon G
TG
Suh
Pann-Ghill
PG
Ryu
Sung Ho
SH
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid / Q000379:methods*; D003720:Densitometry; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives / Q000032:analysis*; D008024:Ligands; D013058:Mass Spectrometry / Q000379:methods*; D036103:Nanotechnology; D010179:Pancreas / Q000737:chemistry; D010447:Peptide Hydrolases / Q000494:pharmacology; D010455:Peptides / Q000032:analysis*; D051381:Rats; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013552:Swine
D007328:Insulin; D008024:Ligands; D010455:Peptides; C063853:insulin, Arg(B31,B32)-; D011972:Receptor, Insulin; D010447:Peptide Hydrolases