A reusable liposome array and its application to assay of growth-hormone-related peptides

Shoji, Atsushi; Sugimoto, Erika; Orita, Sayaka; Nozawa, Keiichiro; Yanagida, Akio; Shibusawa, Yoichi; Sugawara, Masao
June 2010
Analytical & Bioanalytical Chemistry;Jun2010, Vol. 397 Issue 3, p1377
Academic Journal
We describe a reusable liposome array based on the formation of cleavable disulfide cross-links between liposomes and the surface of a glass slip. The N-succinimidyl 3-(2-pyridyldithio)-propionate (SPDP)-modified liposomes encapsulating a pH-sensitive fluorescence dye were immobilized on a 3-mercaptopropyltrimethoxysilane (MTS)-modified glass slip through the formation of disulfide bonds. The regeneration of a used slip was performed by the lysis of immobilized liposomes with Triton X-100 and the cleavage of disulfide bonds by reduction with TCEP, followed by immobilization of SPDP-modified liposomes. The regeneration steps did not affect the fluorescence intensity of re-immobilized liposomes. The liposome array was applied to simultaneous quantification of growth hormone related peptides, i.e., GHRF and somatostatin, in a mixture. After optimizing the assay condition, the method allowed quantification of GHRF and somatostatin in concentration ranges from 0.5 × 10−9 to 0.5 × 10−7 g/mL with detection limits of 2 × 10−10 and 3 × 10−10 g/mL, respectively.


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