Characteristics of a liposome immunoassay on a poly(methyl methacrylate) surface.
Liposome immunoassay (LIA) is based on enzyme immunoassay (EIA) but the detection sensitivity could be significantly enhanced by using antibody-coupled immunoliposomes encapsulating HRP (horse radish peroxidase). Here, we applied LIA to non-porous poly(methyl methacrylate) (PMMA) and polystyrene (PS) surfaces to compare its detection sensitivity with that of EIA, using rabbit IgG (a ligand molecule) and anti-rabbit IgG antibody (a capture molecule) as the model system. LIA developed much stronger color signals than EIA, especially at a lower concentration range (< ca. 1 microg mL(-1)). PMMA showed higher affinity toward rabbit IgG than the PS surface, and the anti-rabbit IgG antibody adsorbed on PMMA was more stable than that on PS. Furthermore, the effects of spot volume and antibody concentration on the signal density were analyzed. The signal density increased as the antibody concentration increased, but it was not significantly affected by the spot volume (2.5-20 microL). In conclusion, LIA on PMMA as a solid support is a very useful, highly sensitive microarray detection system.
Hwang
Sang Youn
SY
Department of Chemical Engineering, Hanyang University, Ansan, South Korea.
Kumada
Yoichi
Y
Seong
Gi Hoon
GH
Choo
Jaebum
J
Katoh
Shigeo
S
Lee
Eun Kyu
EK
D000818:Animals; D002214:Capsules; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis; D008081:Liposomes / Q000737:chemistry*; D046228:Microarray Analysis / Q000379:methods; D008954:Models, Biological; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D011817:Rabbits; D012680:Sensitivity and Specificity
D002214:Capsules; D007074:Immunoglobulin G; D008081:Liposomes; D011137:Polystyrenes; D019904:Polymethyl Methacrylate