Open-sandwich molecular imprinting: making a recognition matrix with antigen-imprinted antibody fragments.
A novel antibody-polymer conjugation method termed open-sandwich molecular imprinting (OS-MIP) has been proposed to produce a specific recognition matrix in the presence of a target antigen. The resultant carboxymethyldextran matrix conjugated with two separate antibody variable region fragments imprinted with the cognate antigen showed higher antigen-binding capacity than non-imprinted ones and was successfully used to sensitively monitor multiple antigen binding/desorption events by a surface plasmon resonance biosensor. Furthermore, when each fragment was labeled with different fluorophores before conjugation, the fluorescence signals of the matrix made by OS-MIP clearly showed an antigen concentration dependent increase in Förster resonance energy transfer between the two dyes. By using a combination of various methods for detecting interaction, OS-MIP will be a useful platform for detecting various targets from small molecules to proteins with high sensitivity and specificity.
Frontier Core-Technology Laboratories, FUJIFILM Corporation, Kaisei-Machi, Ashigarakami-Gun, Kanagawa, Japan.
Koichi
K
Minami
Masaki
M
Ihara
Shou
S
Kuroda
Hirohiko
H
Tsuzuki
Hiroshi
H
Ueda
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D000941:Antigens / Q000276:immunology*; D003911:Dextrans / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D054802:Molecular Imprinting; D009113:Muramidase / Q000276:immunology; D011108:Polymers / Q000737:chemistry; D020349:Surface Plasmon Resonance
D000906:Antibodies; D000941:Antigens; D003911:Dextrans; D007128:Immunoglobulin Fragments; D011108:Polymers; C014392:carboxymethyl dextran; C421864:hen egg lysozyme; D009113:Muramidase