Abstract
An efficient photoimmobilization technique using photoactivated cross linker, N-succinimidyl 4-benzoylbenzoic acid, coupling with a LabVIEW-controlled automatic printing system for multi-protein chip manufacturing was developed in this study. The precoating of a hydrophilic protein layer, ovalbumin (OVA) or bovine serum albumin (BSA) on glass slide and polyethylene terephalate (PET) film to minimize the non-specific adsorption was tested. Moreover, FITC-BSA was photoimmobilized onto the protein pre-coated PET film by exposing to a 4-W Hg lamp (365nm) for 1 hr at room temperature to understand the applicability of the developed techniques on protein chip development. The non-specific adsorption of protein can br significantly decreased from 15 - 25 % without hydrophilic protein layer to 7 - 9 % when 30 mg/ml BSA or OVA was pre-coated on PET film. Also, the structure conformaton of the mouse IgG can be preserved, allowed the detection by monoclonal antibodies. The spot size 50 um can be easily achieved and minimum size of 5 um can alsi be observed when a photomask was used. Moreoverm the glycerol adition for preventing the evaporatiion of protein solution will not interfere the photoimmobolization and multi-protein chip can be prepared by using serial exposing and printing strategy developed in this study.