Quantitative analysis of human serum leptin using a nanoarray protein chip based on single-molecule sandwich immunoassay

Seungah Lee, Shinae Lee, Young Ho Ko, Hyungil Jung, Jung Dong Kim, Joon Myong Song, Jaebum Choo, Seong Kug Eo, Seong Ho Kang

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We report a method for the quantitative analysis of human serum leptin, which is a protein hormone associated with obesity, using a nanoarray protein chip based on a single-molecule sandwich immunoassay. The nanoarray patterning of a biotin-probe with a spot diameter of 150 nm on a self-assembled monolayer functionalized by MPTMS on a glass substrate was successfully accomplished using atomic force microscopy (AFM)-based dip-pen nanolithography (DPN). Unlabeled leptin protein molecules in human serum were detected based on the sandwich fluorescence immunoassay by total internal reflection fluorescence microscopy (TIRFM). The linear regression equation for leptin in the range of 100 zM-400 aM was determined to be y = 456.35x + 80,382 (R = 0.9901). The accuracy and sensitivity of the chip assay were clinically validated by comparing the leptin level in adult serum obtained by this method with those measured using the enzyme-linked immunosorbent assay (ELISA) performed with the same leptin standards and serum samples. In contrast to conventional ELISA techniques, the proposed chip methodology exhibited the advantages of ultra-sensitivity, a smaller sample volume and faster analysis time.

Original languageEnglish
Pages (from-to)608-612
Number of pages5
JournalTalanta
Volume78
Issue number2
DOIs
Publication statusPublished - 2009 Apr 30

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Protein Array Analysis
Leptin
Immunoassay
Molecules
Assays
Serum
Chemical analysis
Immunosorbents
Proteins
Enzyme-Linked Immunosorbent Assay
Immunosorbent Techniques
Nanolithography
Atomic Force Microscopy
Fluorescence microscopy
Self assembled monolayers
Enzymes
Biotin
Fluorescence Microscopy
Linear regression
Glass

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Lee, Seungah ; Lee, Shinae ; Ko, Young Ho ; Jung, Hyungil ; Kim, Jung Dong ; Song, Joon Myong ; Choo, Jaebum ; Eo, Seong Kug ; Kang, Seong Ho. / Quantitative analysis of human serum leptin using a nanoarray protein chip based on single-molecule sandwich immunoassay. In: Talanta. 2009 ; Vol. 78, No. 2. pp. 608-612.
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Quantitative analysis of human serum leptin using a nanoarray protein chip based on single-molecule sandwich immunoassay. / Lee, Seungah; Lee, Shinae; Ko, Young Ho; Jung, Hyungil; Kim, Jung Dong; Song, Joon Myong; Choo, Jaebum; Eo, Seong Kug; Kang, Seong Ho.

In: Talanta, Vol. 78, No. 2, 30.04.2009, p. 608-612.

Research output: Contribution to journalArticle

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