One-pot glyco-affinity precipitation purification for enhanced proteomics: the flexible alignment of solution-phase capture/release and solid-phase separation.
A one-pot affinity precipitation purification of carbohydrate-binding protein was demonstrated by designing thermally responsive glyco-polypeptide polymers, which were synthesized by selective coupling of pendant carbohydrate groups to a recombinant elastin-like triblock protein copolymer (ELP). The thermally driven inverse transition temperature of the ELP-based triblock polymer is maintained upon incorporation of carbohydrate ligands, which was confirmed by differential scanning calorimetry and (1)H NMR spectroscopy experiments. As a test system, lactose derivatized ELP was used to selectively purify a galactose-specific binding lectin through simple temperature-triggered precipitation in a high level of efficiency. Potential opportunities might be provided for enhanced proteomic, cell isolation as well as pathogen detection applications.
Sun
Xue-Long
XL
Departments of Surgery and Biomedical Engineering, Emory University School of Medicine, Atlanta, Georgia 30322, USA. xsun@emory.edu
Haller
Carolyn A
CA
Wu
XiaoYi
X
Conticello
Vincent P
VP
Chaikof
Elliot L
EL
D000595:Amino Acid Sequence; D002152:Calorimetry, Differential Scanning; D002241:Carbohydrates; D011232:Chemical Precipitation; D004549:Elastin / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005690:Galactose / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D006358:Hot Temperature; D007785:Lactose / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*; D040081:Protein Array Analysis; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D000080506:Sialyl Lewis X Antigen; D013696:Temperature
D002241:Carbohydrates; D009844:Oligosaccharides; D010455:Peptides; D011108:Polymers; D011506:Proteins; D000080506:Sialyl Lewis X Antigen; D004549:Elastin; D005947:Glucose; D007785:Lactose; D005690:Galactose