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Info.
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Vol.2 - No.1 (2008.03.20) |
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Title
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Some Key Factors in a Bead-based Fluorescence Immunoassay |
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Authors
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Young Jun Kim1, Hye-Yoon Kim1, Chil Seong Ah1,Moon-Youn Jung1 & Seon-Hee Park1 |
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Institutions
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1IT Convergence and Component Laboratory, Electronic and
Telecommunications Research Institute, Daejeon 305-700, Korea
Correspondence and requests for materials should be addressed
to Y.J. Kim (junkim@etri.re.kr) |
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Abstract
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In an effort to understand some key factors in a bead-based fluorescence immunoassay, a series of sandwich-type immunoassays were carried out on a glass plate using a latex bead embedded with dye molecules. The fluorescence bead functionalized with an amine functionality was conjugated with an anti- prostate specific antigen (anti-PSA) using glutaraldehyde. The glass surface was functionalized with the amine group using 3-aminopropltriethoxysilane(3-APT). The amine functionality on the glass surface was converted to aldehyde usinglutaraldehyde followed by conjugation with the anti-PSA. A series of sandwich-type immunoassays were carried out by incubating the anti-PSA bead with PSA followed by incubating the PSA/bead complex on the anti-PSA glass surface. The dependency of the fluorescence intensity in the immunoassay was observed to increase when the fluorescence bead concentration was increased from 10-3 mg/mL to 101 mg/mL with some signs of saturation around the range of 100 mg/mL to 101 mg/mL. Also, the fluorescence intensity was observed toincrease when the incubation time of the PSA /anti-PSA bead complex on the anti-PSA glass was increased from 5 minutes to 60minutes. The shear force generated during the washing step was observed to affect the bead-based immunoassay.
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Keyword
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Bead-based immunoassay, Dye-embedded particle, Fluorescence bead, Shear force
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PDF File
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