Macromolecular scaffolds for immobilizing small molecule microarrays in label-free detection of protein-ligand interactions on solid support.
We explored two macromolecular scaffolds, bovine serum albumin (BSA) and polyvinyl alcohol (PVA), as chemically complementary platforms for immobilizing small molecule compounds on functionalized glass slides. We conjugated biotin molecules to BSA and amine-derivatized PVA and subsequently immobilized the conjugates on epoxy-functionalized glass slides through reaction of free amine residues on BSA and PVA with surface-bound epoxy groups. We studied binding reactions of such immobilized small molecule targets with solution-phase protein probes using an oblique-incidence reflectivity difference scanning optical microscope. The results showed that both BSA and amine-derivatized PVA were effective and efficient as carriers of small molecules with NHS residues and fluoric residues and for immobilization on epoxy-coated solid surfaces. A significant fraction of the conjugated small molecules retain their innate chemical activity.
Sun
Y S
YS
Department of Physics, University of California, Davis, California 95616, USA.
Landry
J P
JP
Fei
Y Y
YY
Zhu
X D
XD
Luo
J T
JT
Wang
X B
XB
Lam
K S
KS
D000818:Animals; D001710:Biotin / Q000737:chemistry / Q000276:immunology; D002417:Cattle; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D008024:Ligands; D011142:Polyvinyl Alcohol / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D012710:Serum Albumin, Bovine; D001710:Biotin; D011142:Polyvinyl Alcohol