earticle

영어

A new protein chip for fluorescence immunoassay was developed by thermal deposition of modified parylene film on a slide glass for the one-step immobilization of biomolecules. Conventional parylene is a polymer of p-xylene, and it is modified to have aldehyde groups (-CHO) in its molecular structure. The aldehyde groups made covalent bonding with amine group on proteins and peptides by one-step incubation. The efficiency of immobilization and the long-term stability of the modified parylene film were determined to be far improved in comparison to the conventional protein chips by using horseradish peroxidase (HRP) as a model protein. The applicability to the fluorescence immunoassay based on the protein chip was demonstrated by the detection of antibodies against cyclic citrullinated peptide (CCP) in the patient serum. The CCP was known to be an antigne against the autoantibodies of Rheumatoid Arthritis (RA). Therefore, the detection of antibodies agains CCP could be used for the early diagnosis of RA. In this work, CCP was immobilized on the protein chip and the amount of autoantibodies against CCP was quantified by using Cy-5 labeled anti-hIgG antibodies. By comparison with the test results (n=30) from conventional test, the feasibility of RA diagnosis by using the protein chip was confirmed.