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erhaps most significantly are the big increases in assay sensitivity and the widening of the dynamic range accomplished by using alternatives to the traditional colorimetric detection, or by the use of new platform technologies.Several of the new offerings utilise bead or particle-based components some of which are advantageous for multiplexing.This was followed by plasma (48% analysing); cultured cells (41% analysing); whole blood (31% using) and then cell lysate (30% using).
Other automated instrumentation is designed to support the creation/coating of ELISA assay plates or contains cartridge-based delivery of assay reagents to reduce set-up time.
New developments in automated plate washing and the minimisation of wash steps are also impacting conventional ELISAs.
Most respondents (33% each) have either not applied any automation to their ELISA assays today (2012), or ELISA automation has been limited to plate washing only.
Of the remainder, 13% have partially automated ELISA (ie sample/reagent additions, plate washing and incubation cycles); 12% have applied fully automated robotic processing to non-homogeneous ELISA (ie sample/reagent additions, plate washing, incubation and detection); 1% have partially automated ELISA by a homogeneous method (ie sample/reagent additions and incubation) and so far no respondents have applied fully automated robotic processing (ie including detection) to homogeneous ELISA (Figure 4).
ELISA has gained widespread acceptance across a very diverse range of fields (eg diagnosis of infectious diseases, food allergen detection, plant pathogens to biomarkers), but it never achieved significant adoption by early drug discovery labs.
This initially reflected the fact that plate washing was perceived to be difficult to automate, and subsequently the industry’s preference for homogenous screening methods, that could be miniaturised.
For most of this time a heterogeneous, solid-phase microplate-based format utilising multiple washing steps and a colorimetric readouts has predominated.
However, numerous variants of this format have been developed over the years (eg indirect, sandwich, competitive, etc) and a multitude of detection chemistries have been applied to ELISA, including chemiluminescence and various fluorescent readouts.
The enzyme/detection chemistries most used by survey respondents today (2012) in their ELISA assays was HRP (horseradish peroxidase)-colorimetric (88% using); followed by AP (alkaline phosphatase)- colorimetric (31% using) and then HRPchemiluminescent (29% using).
In contrast, only 13% were using electrochemiluminescent detection (Mesoscale Discovery) and 8% using Alpha Screen/Alpha LISA (Perkin Elmer) (Figure 3).