Giant unilamellar vesicles containing Rhodamine 6G as a marker for immunoassay of bovine serum albumin and lipocalin-2.
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.
Department of Chemistry, College of Humanities and Sciences, Nihon University, Setagaya, Tokyo 156-8550, Japan.
Misato
M
Sakamoto
School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Atsushi
A
Shoji
Department of Chemistry, College of Humanities and Sciences, Nihon University, Setagaya, Tokyo 156-8550, Japan. Electronic address: sugawara@chs.nihon-u.ac.jp.
Masao
M
Sugawara
D000818:Animals; D002417:Cattle; D005456:Fluorescent Dyes / Q000032:analysis; D006801:Humans; D007118:Immunoassay; D000071068:Lipocalin-2 / Q000097:blood*; D012235:Rhodamines / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis*; D053835:Unilamellar Liposomes
D005456:Fluorescent Dyes; D000071068:Lipocalin-2; D012235:Rhodamines; D053835:Unilamellar Liposomes; C026188:rhodamine 6G; D012710:Serum Albumin, Bovine
BSA; Giant unilamellar vesicles; Lipocalin-2; Rhodamine 6G; Sandwich-type immunoassay