Journal
ANALYTICAL BIOCHEMISTRY
Volume 505, Issue -, Pages 66-72Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.04.011
Keywords
Giant unilamellar vesicles; Sandwich-type immunoassay; Rhodamine 6G; Lipocalin-2; BSA
Funding
- Ministry of Education, Culture, Sports, Science, and Technology of Japan [21350045]
- College of Humanities and Sciences at Nihon University
- Grants-in-Aid for Scientific Research [25410149] Funding Source: KAKEN
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Functionalized giant unilamellar vesicles (GUVs) containing a fluorescence dye Rhodamine 6G is proposed as a marker in sandwich-type immunoassay for bovine serum albumin (BSA) and lipocalin-2 (LCN2). The GUVs were prepared by the electroformation method and functionalized with anti-BSA antibody and anti-LCN2 antibody, respectively. The purification of antibody-modified GUVs was achieved by conventional centrifugation and a washing step in a flow system. To antigen on an antibody slip, antibody-modified GUVs were added as a marker and incubated. After wash-out of excess reagents and lysis of the bound GUVs with Triton X-100, the fluorescence image was captured. The fluorometric immunoassays for BSA and LCN2 exhibited lower detection limits of 4 and 80 fg ml(-1), respectively. (C) 2016 Elsevier Inc. All rights reserved.
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