mu FBI : A Microfluidic Bead-Based Immunoassay for Multiplexed Detection of Proteins from a mu L Sample Volume

• Background: Over the last ten years, miniaturized multiplexed immunoassays have become robust, reliable research tools that enable researchers to simultaneously determine a multitude of parameters. Among the numerous analytical protein arrays available, bead-based assay systems have evolved into a key technology that enables the quantitative protein profiling of biological samples whilst requiring only a minimal amount of sample material. Methodology/Principal Findings: A microfluidic bead-based immunoassay, mu FBI, was developed to perform bead-based multiplexed sandwich immunoassays in a capillary. This setup allows the simultaneous detection of several parameters and only requires 200 ng of tissue lysate in a 1 mu L assay volume. In addition, only 1 mu L of detection antibodies and 1 mu L of the reporter molecule Streptavidin-Phycoerythrin were required. The mu FBI was used to compare the expression of seven receptor tyrosine kinases and their degree of tyrosine phosphorylation in breast cancer tissue and in normal tissue lysates. The total amount of HER-2, as well the degree of tyrosine phosphorylation was much higher in breast cancer tissue than in normal tissue. mu FBI and a standard bead-based assay led to identical protein expression data. Moreover, it was possible to reduce the quantity of sample material required by a factor of 100 and the quantity of reagents by a factor of 30. Conclusions/Significance: The mu FBI, microfluidic bead-based immunoassay, allows the analysis of multiple parameters from a very small amount of sample material, such as tumor biopsies or tissue sections.

Ämnesord

NATURVETENSKAP  -- Kemi -- Analytisk kemi (hsv//swe)

NATURAL SCIENCES  -- Chemical Sciences -- Analytical Chemistry (hsv//eng)

Nyckelord

BREAST-CANCER

MICROARRAY ANALYSIS

IMMUNOAFFINITY CE

PHOSPHORYLATION

BIOPSIES

ARRAYS

CELLS

TECHNOLOGY

ACTIVATION

PREDICTION

Analytical chemistry

Analytisk kemi

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)