Earticle

초록

영어

Protein microarrays are studied as a rapid analysis of multiple samples, and a critical point for an optical detection signal is ligand density on a chip surface[1]. In this study, to evaluate the activity of the adsorbed ligand proteins on the hydrophilic or hydrophobic surface. We immobilized ligand proteins on different membranes: NC (nitrocellulose) membrane as a hydrophilic surface and PVDF (polyvinylidenedifluoride) membrane as a hydrophobic surface. We used rabbit IgG as the ligand for direct ELISA, and anti-rabbit IgG antibody as the ligand for sandwich ELISA. The adsorption of anti-rabbit IgG antibody to the membrane required 12 hr at 4℃. In sandwich ELISA, anti-rabbit IgG antibody with HRP (horseradish peroxidase) could specifically bind to rabbit IgG antibody. We obtained the results of ligand antibody adsorption and their activity and specificity toward antigen binding on each surface for comparison. In the case of NC membrane, diffusion of a spot was shown faster and wide. Resulting in a lower surface density than PVDF membrane. In conclusion, the optimal surface density was saturated in 10 uL volume and concentration of 2.5 ug/mL of anti-rabbit IgG antibody. Additionally, we apply this experiment to glass surface, too. We compared conjugation with adsorption of immobilized protein on the glass surface. In the case of glass surface, conjugation of proteins was essential process. Because adsorption of proteins was washed out after washing buffer[2]. So we have to use conjugation method of ligand proteins for microarray chip. And we confirmed the possibility of a new microarray system.

저자

• Chang Hwan Lim [ Department of Bio-nanotechnology Han yang University ]
• Eun Kyu Lee [ Department of Bio-nanotechnology Han yang University ]

참고문헌

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간행물

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• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576