The boundary molecules in a lysozyme pattern exhibit preferential antibody binding.
Lysozyme was immobilized on a prefabricated carboxylic acid terminated chemical template, forming a tightly packed, one monolayer thick lysozyme pattern. Polyclonal anti-lysozyme antibodies can bind to the immobilized lysozyme pattern. Atomic force microscope (AFM) observation reveals that the antibodies bind to the lysozyme molecules on the pattern edge before they bind to the lysozyme molecules in the pattern interior. Better spatial accessibility and flexibility of the lysozyme molecules on the pattern edge are used to explain the observed antibody binding preference. The topographies of the lysozyme pattern also affect the antibody binding. The antibodies bind to the edge lysozyme from the top if the lysozyme pattern is half-buried in a 10 A deep channel, whereas the antibodies bind to the edge lysozyme from the side if the lysozyme pattern is immobilized on a protruding terrace. The observed "edge effect" suggests that, for the same protein coverage, reducing the protein pattern feature to the nanoscale will improve the overall binding activity of the immobilized protein toward the antibody.
Gao
Pei
P
Department of Chemistry, University of Kentucky, Rose Street, Lexington, KY 40506, USA.
Cai
Yuguang
Y
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D000936:Antigen-Antibody Complex / Q000737:chemistry; D002264:Carboxylic Acids / Q000737:chemistry; D002645:Chickens; D004529:Egg White / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force / Q000379:methods; D009113:Muramidase / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D012821:Silanes / Q000737:chemistry; D013055:Spectrophotometry, Infrared / Q000379:methods; D013499:Surface Properties
D000906:Antibodies; D000936:Antigen-Antibody Complex; D002264:Carboxylic Acids; D011506:Proteins; D012821:Silanes; C061189:octadecyltrichlorosilane; D009113:Muramidase