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 (365 nm) for 1 hr at room temperature to understand the applicability of the developed techniques on protein chip development. The non-specific adsorption of proteins can be 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 structural conformation of the mouse IgG can be preserved, allowed the detection by monoclonal antibodies The spot size of 50 µm can be easily achieved and minimum size of 5 µm can also be observed when a photomask was used. Moreover, the glycerol addition for preventing the evaporation of protein solution will not interfere the photoimmobilization and multi-protein chip can be prepared by using serial exposing and printing strategy developed in this study.