Current and Emerging Applications of ELISAs
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Last Updated: May 14, 2024
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Published: May 1, 2024
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Kate Harrison, PhD
Senior Science Writer
Kate Harrison is a senior science writer and is responsible for the creation of custom-written projects. She holds a PhD in virology from the University of Edinburgh. Before working at Technology Networks, she was involved in developing vaccines for neglected tropical diseases, and held a lectureship position teaching immunology.
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Enzyme-linked immunosorbent assays (ELISAs) are a cornerstone of clinical and research laboratories. ELISAs can detect and quantify the presence of antibodies, antigens or proteins in an enormous range of biological and environmental samples.
Since their development in 1971, ELISAs have been adapted for use in a wide variety of fields for numerous applications, from epidemiological monitoring to environmental pollutant analysis. Research into novel biomarkers and ELISA-based technologies continues to improve sensitivity and broaden the potential uses.
Download this listicle to explore the uses of ELISAs in:
- Clinical diagnostics and epidemiology
- Biopharmaceutical and vaccine development
- Food safety analysis
- Environmental analysis
1
Current and Emerging Applications
of ELISAs
Kate Harrison, PhD
Enzyme-linked immunosorbent assays (ELISAs) are a cornerstone of clinical and research laboratories. ELISAs
can detect and quantify the presence of antibodies, antigens or proteins in an enormous range of biological and
environmental samples. Generally carried out in 96-well plates, ELISAs rely on the immobilization of an antigen
or antibody to a solid surface (i.e., the base of the plate) and the subsequent binding of a corresponding antibody
or antigen in a specific manner. Although the most basic form of an ELISA simply detects the presence of a
particular antigen or antibody, quantification can be achieved by adding an antibody conjugated to a substrate
that produces a measurable product, such as fluorescence or a color change.
ELISAs were first developed in 1971, by two independent groups. Both of these simultaneously developed assays
were based on enzymatic reporters, rather than the radioactive reporters used by previous immunoassay
iterations.1 These enzyme-based immunoassays were safer and easier to perform than radioactivity-based
immune assays, and quickly became widespread. Since then, ELISAs have been adapted for use in a wide variety
of fields for numerous applications, from epidemiological monitoring to environmental pollutant analysis. There
are now hundreds of commercially available ELISA kits for the detection and quantification of antibodies and
antigens. However, research continues into the development of novel biomarkers and ELISA-based technologies
to improve sensitivity and broaden the potential uses. Some of these current and emerging applications and
methodologies for ELISAs are discussed below.
Clinical diagnostics and epidemiology
One of the most widespread applications of ELISAs is in clinical diagnostics and epidemiology. ELISAs can
be used to detect both antigen markers of a disease and the antibodies against it, depending on the type of
assay selected. Therefore, they can be used to diagnose an active infection and track the spread of a disease
in an outbreak scenario, and also identify previously infected individuals. To help monitor disease epidemics
epidemiologically, a portable field version of an ELISA would be advantageous for onsite diagnosis of infectious
disease. However, although several avenues of research have been investigated to this end – such as a batteryoperated
ELISA system2 and smartphone-linked ELISA systems – nothing is, as yet, commercially available.3,4
In addition to infectious diseases, ELISAs can also be used to indicate the presence of auto-reactive antibodies.
Both the general presence of autoantibodies, using a plate with a variety of self-antigens adsorbed to the surface,
or autoantibodies related to specific autoimmune diseases can be analyzed, making ELISAs indispensable in
autoimmune disease diagnostics.5 One such example is the identification of rheumatoid factors (RFs). RFs are
antibodies that bind to other antibodies and are commonly found in systemic autoimmune diseases. RFs have
diagnostic and prognostic value in several autoimmune diseases and are part of the specific diagnostic criteria
for rheumatoid arthritis.6 For other autoimmune diseases, like autoimmune connective tissue diseases (CTDs),
ELISAs for autoreactive antinuclear antibodies (ANAs) are emerging as a more sensitive, more specific and less
time-consuming alternative to the current gold standard: indirect immunofluorescence assays.7
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Despite their range and versatility, traditional ELISAs are limited in scenarios requiring extremely high sensitivity
for nano-molar levels of sample, making them unsuitable for early diagnosis or screening of diseases with subtle
physiological changes such as Alzheimer’s disease (AD). However, the recent development of nano-ELISAs is now
opening more avenues for highly sensitive detection. In a nano-ELISA, a detector antibody and a signaling molecule
such as horseradish peroxidase (HRP) are coupled with gold nanoparticles. These nanoparticles act as signal
amplifiers, significantly increasing the detection limit and allowing the detection of antigens or antibodies even in
the picogram range.8 This method has been used to identify the AD biomarker Aβ42 in serum samples at extremely
low levels.9 As cerebral spinal fluid is currently collected for detection of AD biomarkers, this suggests future
potential for nano-ELISAs as a novel, less invasive method for AD diagnosis.10
Biopharmaceutical and vaccine development
Biopharmaceuticals are therapeutics derived from, or created in, biological sources. Demand for these
therapeutics continues to grow as they revolutionize the treatment of a wide range of diseases, including cancer
and autoimmune diseases. As they are produced from biological sources, they must be purified to an extremely
high standard to remove any host cell proteins (HCPs) or residual manufacturing reagents. ELISAs are one of the
standard tools for quantifying the total levels of residual impurities, due to their high-throughput, high sensitivity
and high specificity.11 However, they are unable to give any information on the specific, individual HCPs present, so
are now often coupled with liquid chromatography-mass spectrometry (LC-MS) for more in-depth evaluation of
biopharmaceutical purity.
One of the simplest uses for ELISAs is the quantification of antibodies in biological samples. This is essential in
the development of novel vaccines. Many vaccines protect against infection by inducing the creation of specific
antibodies, meaning quantification of antibody levels post-vaccination is needed to assess vaccine efficacy. ELISAs
have therefore been used extensively in the clinical development of approved vaccines, including the ChAdOx1
nCoV-19 vaccine against SARS-CoV-2 and the recently approved R21 and RTS,S vaccines against malaria.12,13,14
Forensic investigations
One of the most common applications for ELISAs in forensic science is drug testing. Toxicologists can test forensic
samples for a wide range of illicit substances, including cocaine, amphetamines, opiates, cannabinoids and
benzodiazepines. Both liquid samples, such as blood and urine, and solid samples such as hair can be processed
for ELISA testing, for purposes such as monitoring drug abstinence and workplace testing.15,16 ELISAs can also be
used to determine if a blood sample is human for forensic investigation by using antibodies targeting specifically
human blood components.15 By targeting the assay at highly abundant antigens in the blood, such as human
albumin, blood can still be identified as human by ELISA even in ancient, buried samples over 3,000 years old.17
In addition to analyzing bodily fluids, ELISAs can also be used in forensic investigations to help determine a cause
of death. Traumatic brain injury (TBI) is one of the leading causes of mortality worldwide and is responsible for
approximately 30% of all injury-related deaths.18 Post-mortem identification of TBI can be difficult, as ante-mortem
clinical diagnosis usually involves neurological assessment. Recent research has shown elevated levels of cerebral
protein biomarkers, such as tau protein and myelin basic protein, in blood and cerebral spinal fluid samples from
cases where TBI was the confirmed cause of death. These markers can therefore be used as an indicator of TBI in
post-mortem examination, even in the absence of visible central nervous system damage.19,20,21
Food safety
ELISAs play a significant role in food safety testing and are used to screen for contaminants and allergens.
Food allergies affect approximately 2% of the Western population, and allergic and anaphylactic reactions
CURRENT AND EMERGING APPLICATIONS OF ELISAS 3
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to food result in 30,000 hospital visits and 150 deaths per year in the United States alone.22 Allergies can
be extremely sensitive. For example, even trace amounts of peanuts can trigger an anaphylactic reaction.
Therefore, it is very important that foods labeled as allergen-free, are indeed safe for consumption.
Cross-contact with allergens can occur at many stages of food processing and production and is often
the result of failure to clean machines appropriately when switching between allergen-containing and
allergen-free foods. However, ELISAs are used by manufacturers to detect common allergens in food
products, including nuts, meat, milk and milk products, fish and soybeans. The highly sensitive and specific
nature of ELISAs can identify cross-contact, preventing the need for food recalls and protecting allergic
populations. However, as large numbers of batch samples will typically need to be tested on a regular
basis, ELISAs, despite their relative speed, can still result in bottlenecks. Novel, rapid microfluidic-based
ELISA platforms are currently being researched to develop a method for more sensitive and far faster
allergen identification.23,24
Despite their widespread use in the field, traditional ELISA techniques can lack the sensitivity needed to
detect extremely low, but still dangerous, levels of toxic or microbial contaminants. However, the development
of nano-ELISAs have enabled the detection of harmful microbes, pesticides, drug residues and
pollutants in food.25 Nano-ELISA techniques have now been developed to identify a range of food contaminants,
including aflatoxin in corn, pathogenic Listeria monocytogenes in milk and veterinary drug traces in
poultry.26,27,28
Environmental analysis
Although ELISAs have typically been used for biological sample analysis, their applications in environmental
sampling is increasing, particularly for the detection and quantification of agricultural and industrial
pollutants in water. The use of pesticides has increased steadily over the last 30 years worldwide,
resulting in increased levels of pollution and potentially toxic agricultural runoff.29 Often, analysis of
environmental soil and water samples for pollutants uses techniques such as gas and liquid chromatography
(GC and LC). However, these techniques can be costly and time-consuming compared to ELISAs,
which have been shown to be just as sensitive as chromatography methods.30 Indeed, ELISAs, along with
other immune-based assays, have been used successfully in several large-scale water quality surveys in
the US.30
Pharmaceutical pollutants, such as hormones and drugs, can also be assessed in both water samples
and the tissues of aquatic organisms by ELISA.31 These emerging pollutants can have a significant effect
on aquatic ecosystems, damaging animal life, altering microbial populations leading to detrimental algal
blooms and perpetuating antibiotic resistance. Monitoring these pollutants is essential for minimizing
harm and developing better regulatory guidelines to prevent their release into the environment.32
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CURRENT AND EMERGING APPLICATIONS OF ELISAS 4
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About the author:
Kate Harrison is a senior science writer and is responsible for the creation of custom-written projects.
She holds a PhD in virology from the University of Edinburgh. Before working at Technology Networks,
she was involved in developing vaccines for neglected tropical diseases and held a lectureship position
teaching immunology.
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