High-throughput immunophenotyping by surface plasmon resonance imaging

Koichi Kato, Toshinari Ishimuro, Yusuke Arima, Isao Hirata, Hiroo Iwata

研究成果: ジャーナルへの寄稿学術誌査読

21 被引用数 (Scopus)

抄録

Cell binding assays on antibody arrays permit the rapid immunophenotyping of living cells. The throughput of the analysis, however, is still limited due to our inability to perform parallel and quantitative detection of cells captured on the array. To address this limitation, we employed here an imaging technique based on surface plasmon resonance (SPR). SPR has been frequently used to monitor capture of proteins on antibody microarrays, while few cases were reported for capture of cells. Antibody arrays were prepared through the photopatterning of an alkanethiol monolayer on a gold-evaporated glass plate and the subsequent immobilization of various antibodies onto 4-9 separate spots created by photopatterning. A glass slip was mounted onto the array with a thin spacer to construct a parallel-plate chamber. Leukemia cells were injected into the chamber to conduct a binding assay, while refractive index changes at the vicinity of the array surface were monitored by SPR imaging. We observed that SPR signals were intensified on specific antibody spots but not on nonspecific spots. Confocal laser scanning microscopy revealed that the observed SPR signals were attributed to cell deformations caused by multivalent interactions with immobilized antibody, which effectively elevated the refractive index of a medium phase within an evanescent field. This effect could be suitably utilized to monitor quantitatively cell binding to multiple spots from a heterogeneous cell population.

本文言語英語
ページ(範囲)8616-8623
ページ数8
ジャーナルAnalytical chemistry
79
22
DOI
出版ステータス出版済み - 11月 15 2007
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 分析化学

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