Liposome-based immunoaffinity chromatographic assay for the quantitation of immunoglobulin E in human serum.
Immunoglobulin E (IgE)-mediated type I allergies affect over 25% of the world's population; they are among the most common diseases in developed countries. Therefore, simple and rapid in vivo and in vitro methods for diagnosing allergies are becoming increasingly important. In this paper, we demonstrate the feasibility of using sulforhodamine B, a fluorescent dye, entrapped inside immunoliposomes, the outer surfaces of which were sensitized with IgE, as a signal amplifier for the development of a simple, rapid, and inexpensive colorimetric affinity chromatographic immunoassay for the detection of total IgE in serum. This assay operates based on competition between standards (or human serum samples) containing IgE and IgE-sensitized immunoliposomes for the limited number of antigen binding sites of immobilized anti-IgE antibodies at the antigen capture (AC) zone on the nitrocellulose membranes. The color density of the AC zone is indirectly proportional to the number of IgE units present in the test sample. The detection limit of this liposome-based immunoaffinity chromatographic assay was 0.37ng in IgE-free serum solution (equivalent to 20microL of a 18.5ngmL(-1) solution). A commercially available ELISA kit was used as a reference method to validate the proposed assay through the analysis of three human serum samples.
Annie Ho
Ja-an
JA
Department of Chemistry, National Tsing Hua University, Hsinchu, Taiwan. jaho@mx.nthu.edu.tw
Wu
Li-Chen
LC
Chang
Li-Hui
LH
Hwang
Kuo-Chu
KC
Reuben Hwu
Jih-Ru
JR
D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood*; D007163:Immunosorbent Techniques; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D015203:Reproducibility of Results; D012235:Rhodamines / Q000737:chemistry; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D008081:Liposomes; D012235:Rhodamines; C022027:lissamine rhodamine B; D007073:Immunoglobulin E