Oriented immobilization of antibodies by a self-assembled monolayer of 2-(biotinamido)ethanethiol for immunoarray preparation.
Orientation of antibodies is very important in the preparation of immunoarrays to keep the activity of antibodies on solid surfaces. Thus, we synthesized a new bifunctional compound, 2-(biotinamido)ethanethiol, and investigated whether the thiol compound is useful to analyze antibody-antigen interactions on immunoarrays with a spectral SPR biosensor. The synthesized organic thiol was characterized by nuclear magnetic resonance spectroscopy and mass spectrometry. 2-(Biotinamido)ethanethiol formed a monolayer on a gold surface and properly immobilized antibodies via streptavidin and biotinylated protein G. Optimal molar ratio of 2-(biotinamido)ethanethiol and mercaptohexanol for antigen-antibody interactions was 1:2. Thus, 2-(biotinamido)ethanethiol is an useful bifunctional linker in the preparation of immunoarrays on gold surfaces.
Jung
Se-Hui
SH
Department of Molecular and Cellular Biochemistry and Nano-Bio Sensor Research Center, Kangwon National University College of Medicine, and Chunchon, Kangwon-Do 200-701, South Korea.
Son
Ha-Young
HY
Yuk
Jong Seol
JS
Jung
Jae-Wan
JW
Kim
Kyung Hwan
KH
Lee
Chang-Hee
CH
Hwang
Hoon
H
Ha
Kwon-Soo
KS
D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D019921:Biotinylation; D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry; D006046:Gold / Q000737:chemistry; D000441:Hexanols / Q000378:metabolism; D007118:Immunoassay; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D015394:Molecular Structure; D009419:Nerve Tissue Proteins / Q000378:metabolism; D040081:Protein Array Analysis; D019809:Streptavidin / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000378:metabolism
C509499:2-(biotinamido)ethanethiol; D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D020099:Coated Materials, Biocompatible; D003432:Cross-Linking Reagents; C029152:G-substrate; D000441:Hexanols; D009419:Nerve Tissue Proteins; D013438:Sulfhydryl Compounds; D003543:Cysteamine; D001710:Biotin; D006046:Gold; D019809:Streptavidin; C477019:3-mercaptohexanol