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Most (36%) of survey respondents were using a 96-well whole plate washer for their ELISA assays.The next most used washer type was an 8 or 12- channel strip washer (30% using), followed by manual/hand washed assays (22% using).The majority (57%) of survey respondents do not see the need for commercially available ELISA kits to be miniaturised, indicating that the standard volume 96-well format meets their current needs.
This review article uses as its starting point some of the findings of HTStec’s ELISA Assay Trends 2012 survey and report, published in June 2012 (1).
This report discusses end-users’ current practices and preferences in ELISA assays, and seeks to understand future user requirements.
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 preferred manufacturer of these ELISA washers was Bio Tek (Figure 5).
The majority (51%) of survey respondents were using the 96-well microplate as their primary assay format for ELISA assays today (2012).These are supported by a wide range of ELISA reagents, assay and kits providing relative profiling or absolute quantitation of proteins across a wide range of study areas.When coupled with the breadth of existing global expertise in immunoassays, it is easy to see why ELISA still remains the methodology of choice by those labs tasked with analyte detection.Among those responding no interest was found for blood banking, stem cells, environmental or quality control as the main (primary) applications of ELISA assays (Figure 1).Of the sample source analysed by ELISA most respondents (67%) were analysing serum.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.The immunoassay landscape has, however, dramatically changed in recent years with the development of reduced assay steps protocols, no wash ELISAs and new automated systems designed to run ELISA.More recently, bead-based methods and microfluidic platforms have started to impact immunoassays and by coupling with single molecule detection look set to take the sensitivity of traditional ELISA to new levels.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).