Microfluidic Analysis of Antibody Specificity in a Compact Disk Format

A new and flexible technology for high throughput analysis of antibody specificity and affinity is presented. The method is based on microfluidics and takes advantage of compact disks (CDs) in which the centrifugal force moves fluids through microstructures containing immobilized metal affinity chro...

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Bibliographic Details
Published inJournal of proteome research Vol. 5; no. 7; pp. 1568 - 1574
Main Authors Eriksson, Cecilia, Agaton, Charlotta, Kånge, Rikard, Sundberg, Mårten, Nilsson, Peter, Ek, Bo, Uhlén, Mathias, Gustafsson, Magnus, Hober, Sophia
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 01.07.2006
Subjects
Animals
Antibodies - analysis
Antibody Specificity
biochip
Bioengineering
Bioteknik
CHIP
Chromatography, Affinity
compact disk
Compact Disks
GENOME
gyrolab
IMMUNOSORBENT-ASSAY ELISA
MICROARRAYS
Microfluidic Analytical Techniques - instrumentation
Microfluidic Analytical Techniques - methods
microfluidics
Miniaturization
Protein Array Analysis
PROTEOMICS
Rabbits
Reproducibility of Results
Sensitivity and Specificity
TECHNOLOGY
TEKNIKVETENSKAP
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Summary:A new and flexible technology for high throughput analysis of antibody specificity and affinity is presented. The method is based on microfluidics and takes advantage of compact disks (CDs) in which the centrifugal force moves fluids through microstructures containing immobilized metal affinity chromatography columns. Analyses are performed as a sandwich assay, where antigen is captured to the column via a genetically attached His6-tag. The antibodies to be analyzed are applied onto the columns. Thereafter, fluorescently labeled secondary antibodies recognize the bound primary antibodies, and detection is carried out by laser-induced fluorescence. The CDs contain 104 microstructures enabling analysis of antibodies against more than 100 different proteins using a single CD. Importantly, through the three-dimensional visualization of the binding patterns in a column it is possible to separate high affinity from low affinity binding. The method presented here is shown to be very sensitive, flexible and reproducible. Keywords: microfluidics • miniaturization • compact disk • biochip • gyrolab • antibody specificity
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ISSN:1535-3893
1535-3907
1535-3907
DOI:10.1021/pr050447c