Biomarkers
practical complexities of large multiplex assays. Among the various multiplex methods, the Luminex xMAP® platform was generally reported to have better precision than the Cytometric Bead Array13 or electrochemiluminescence-based Meso- Scale systems14.
These multiplex technologies require the use of dedicated instruments with limited flexibility for additional analytical applications. Multiplex assays also require complex mixtures of numer- ous antibodies against a wide variety of biomark- ers in the same sample. Although validated panels are generally available for broad applications, these panels might not contain specific targets of interest, especially if an unbiased selection strate- gy was employed for biomarker identification. In addition, assay performance may not be optimal for each analyte, restricting the lower limit of detection for some targets in the panel. For these reasons, multiplex assays are often used to get a larger picture of biomarker modulation which is then used to select a smaller subset of more rele- vant targets.
The Alpha Technology platform from PerkinElmer (eg AlphaLISA® and AlphaScreen®) represents a homogenous no-wash ELISA assay alternative that allows samples to be analysed effi- ciently without compromising sensitivity in com- plex biological matrices such as serum, plasma, cell culture supernatants or crude cell lysates. This assay platform is less time-intensive than tradition- al ELISA, allowing for simple ‘add-and-read’ pro- tocols that enable automated high-throughput applications for rapid biomarker validation. Alpha assays are also truly miniaturisable, a crucial fea- ture when quantitative detection of analytes must be performed on precious biological samples avail- able only in limited supply. AlphaLISA assays are highly robust when using sample volumes as low as 1µl in total assay volumes of 10µl. Validated AlphaLISA kits are available to study close to 100 major biomarkers in inflammation, cancer, cardiovascular, Alzheimer’s or metabolic diseases. AlphaLISA is a bead-based proximity assay, whereby the target is recognised by specific antibodies on two different beads (Figure 2).
Address Even the Most Di cult Assays
RapidFire®enables label-free detec on of na ve analytes for previously intractable targets
Label-free screening for:
• oncology • epigene cs • metabolic disorders • cardiovascular disease • an -infec ves • neurological disorders • in amma on
T: 781.721.3695 | F: 781.998.0054 |
www.BIOCIUS.com |
info@biocius.com Drug Discovery World Summer 2010 19
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