Preparation of a protein micro-array using a photo-reactive polymer for a cell-adhesion assay.
A protein micro-array, called a "cell chip" was constructed by using a photo-reactive polymer for a cell-adhesion assay. Various amounts of albumin or fibronectin were covalently immobilized on a polystyrene dish using a micro-spotter with a dip pen. First, poly(acrylic acid) carrying azidophenyl groups was synthesized as the photo-reactive polymer. Secondly, the aqueous solution of a photo-reactive polymer (several nanoliters) was cast using the dip pen of the micro-spotter and dried in air. Subsequently, aqueous solutions of proteins were cast on the same place using the micro-spotter. After drying, the dish was irradiated with ultraviolet light. Finally, the immobilization was confirmed by staining with a dye. The immobilization was stable even after washing with Tween-20. The protein-immobilized area depended on the manipulation of the micro-spotter and the size of the dip pen. Subsequently, cell adhesion on the photo-immobilized protein micro-array was investigated. The adhesion behavior of cells depended on the kind of immobilized proteins and the kind of cells. The protein micro-array will be useful for cell diagnosis and for the selection of biomaterials to regulate cell behavior.
Ito
Yoshihiro
Y
Kanagawa Academy of Science and Technology, KSP East 309, Sakado 3-2-1, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
Nogawa
Masayuki
M
D000180:Acrylic Resins; D000818:Animals; D019556:COS Cells; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002460:Cell Line; D002522:Chlorocebus aethiops; D020099:Coated Materials, Biocompatible / Q000494:pharmacology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005353:Fibronectins / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D016716:PC12 Cells; D010777:Photochemistry / Q000295:instrumentation / Q000379:methods; D011108:Polymers / Q000528:radiation effects; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D012710:Serum Albumin, Bovine / Q000494:pharmacology*
D000180:Acrylic Resins; D020099:Coated Materials, Biocompatible; D005353:Fibronectins; D011108:Polymers; D012710:Serum Albumin, Bovine; C006903:carbopol 940