Immobilized peptides as high-affinity capture agents for self-associating proteins.
There is currently great interest in the fabrication of protein-detecting arrays comprised of large numbers of immobilized protein capture agents. While most efforts in this arena have focused on the use of biomolecules such as antibodies and nucleic acid aptamers as capture agents, synthetic species have many potential advantages. However, synthetic molecules isolated from combinatorial libraries generally do not bind target proteins with the high affinity necessary for array applications. Here, we demonstrate that simple linear peptides bind dimeric proteins tenaciously when immobilized, although they exhibit only modest affinity in solution. These data show that high-affinity bidentate capture agents for dimeric proteins can be created by simply immobilizing modest-affinity ligands on a surface at high density, bypassing the requirement for careful optimization of linker length and geometry that is normally required to create a high-affinity solution bidentate ligand.
Naffin
Jacqueline L
JL
Department of Internal Medicine, Center for Biomedical Inventions, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
Han
Ying
Y
Olivos
Hernando J
HJ
Reddy
M Muralidhar
MM
Sun
Tingwan
T
Kodadek
Thomas
T
D020650:Combinatorial Chemistry Techniques; D019281:Dimerization; D004355:Drug Stability; D005453:Fluorescence; D008024:Ligands; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D011506:Proteins