Signal amplification of microarray-based immunoassay by optimization of nanoliposome formulations.
The use of microarray-based immunoassay is often limited by its sensitivity. To increase the sensitivities of such an immunoassay, liposome encapsulation was explored. Two different liposome formations and several preparation methods were examined to optimize encapsulation and signal-enhancing efficacy for enzyme-linked immunosorbent assay (ELISA) and antibody array. The signal amplification by liposome encapsulation was demonstrated through a detection for foodborne pathogenic Listeria. In plate-trapped antigen (PTA) ELISA, horseradish peroxidase (HRP)-loaded liposome increased signal 9-fold more than the control. Limits of detection (LODs) of HRP-encapsulated liposome were 6.4 × 10(5) and 5.5 × 10(6)CFU/ml in sandwich ELISA and antibody array, respectively. Furthermore, when chromogenic 4-chloro-1-naphthol (4-CN) substrate was used for signal development in the antibody array, the signal could be detected with the naked eye. These results suggest that the liposome encapsulation technique can have great potential for signal amplification and, therefore, for increasing assay sensitivity for various formats of immunoassay, especially microarray-based format.
National Nanotechnology Center (NANOTEC), Pathumthani 12120, Thailand.
Uracha
U
Ruktanonchai
Onanong
O
Nuchuchua
Ratthaphol
R
Charlermroj
Thitiporn
T
Pattarakankul
Nitsara
N
Karoonuthaisiri
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000032:analysis; D002626:Chemistry, Pharmaceutical; D003124:Colorimetry; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D007118:Immunoassay; D008081:Liposomes / Q000737:chemistry*; D008087:Listeria / Q000378:metabolism; D036103:Nanotechnology; D009284:Naphthols / Q000737:chemistry; D040081:Protein Array Analysis
D000906:Antibodies; D000941:Antigens; D008081:Liposomes; D009284:Naphthols; C025917:4-chloro-1-naphthol; D006735:Horseradish Peroxidase