• EBOV
• NiV
• chemokines
• choroid plexus
• cytokines
• neurotropism
• organotypic brain slices

• Animals
• Ebolavirus/physiology
• Ebolavirus/pathogenicity
• Nipah Virus/physiology
• Nipah Virus/pathogenicity
• Mice
• Brain/virology
• Brain/pathology
• Cytokines/metabolism
• Viral Tropism
• Hemorrhagic Fever, Ebola/virology
• Hemorrhagic Fever, Ebola/pathology
• Virus Replication
• Henipavirus Infections/virology
• Mice, Inbred C57BL
• Disease Models, Animal
• Humans
• Mice, Knockout
• Receptor, Interferon alpha-beta/genetics
• Receptor, Interferon alpha-beta/metabolism

• IgY
• agglutinin
• antinutritional factor
• haemagglutination
• soybean

• Animals
• Rocky Mountain Spotted Fever/diagnosis
• Rocky Mountain Spotted Fever/microbiology
• Biosensing Techniques
• Immunoassay
• Rodentia/microbiology
• Immunoglobulin G

• IgY
• antibaceterial
• antigens
• avian antibodies (IgY)
• hyperimmune
• infection

• Politechnika Wrocławska

• Anti-phospholipase A2
• Antioxidant capacity
• Lipid oxidation
• Lipidomics
• Phospholipase A2
• Phospholipid

• IgY
• calicivirus
• foodborne disease
• gastroenteritis outbreaks
• norovirus
• outbreak prevention and control
• passive immunotherapy

• Humans
• Animals
• Female
• Chickens
• Feasibility Studies
• Caliciviridae Infections/prevention & control
• Caliciviridae Infections/veterinary
• Norovirus/genetics
• Blood Group Antigens/genetics
• Antibodies

This research aims to evaluate the feasibility of using avian immunoglobulins (IgY) raised against adhesion factors of enterotoxigenic Escherichia coli (ETEC) as prophylaxis of diarrheal illness caused by these pathogens. ETEC requires adhesion to human intestinal epithelial cells as a primary step in establishing enteric infection. Therefore, inhibition of adhesion may prevent such infections and reduce clinical burdens of diarrheal illness.

IgY samples were prepared from eggs of hens immunized with an adhesin-tip multiepitope fusion antigen (MEFA), developed against nine adhesin tip epitopes derived from clinically relevant ETEC strains. The resulting IgY was evaluated for its ability to inhibit adhesion of ETEC to cell-surface targets. Potential impacts of anti-MEFA IgY on growth of both pathogenic and commensal E. coli isolates were also evaluated.

Enzyme linked immunosorbent assay (ELISA) titers were achieved for IgY targeting each of the nine individual epitopes included in the adhesin-tip MEFA. Furthermore, anti-MEFA titers exceeding 1:2¹⁹ were sustained for at least 23 weeks. All ETEC strains used in design of the adhesin-tip MEFA, and five of five clinical ETEC strains were significantly (P < 0.05) inhibited from adhesion to mammalian cells in culture.

These findings demonstrate that IgY targeting ETEC adhesin-tip MEFA have the potential to disrupt in vitro adherence of ETEC. A formulation containing adhesin-tip MEFA IgY can be considered a potential candidate for in vivo evaluation as prophylaxis of diarrheal diseases. Animal studies of this formulation are planned.

• Animals
• Female
• Humans
• Enterotoxigenic Escherichia coli
• Chickens
• Feasibility Studies
• Antigens, Bacterial
• Escherichia coli Infections/prevention & control
• Antibodies, Bacterial
• Adhesins, Bacterial
• Diarrhea/prevention & control
• Epitopes
• Mammals

Hens are oviparous vertebrates and produce IgY antibodies, which is the main type of immunoglobulin in the egg yolk, and high concentrations can be obtained by using a simple method that does not require sophisticated equipment and reagents. The Polyethylene Glycol 6000 method allows the removal of lipids and the precipitation of IgY in two days with an approximated purity of around 80%, however during the original protocol other contaminant proteins can be precipitated. To overcome the issue of contamination with other proteins and extraction time, we optimized the previously method described by Pauly et al. (2011) by adding some changes that improved the aforementioned problems.

• Our protocol is customized by the addition of one more filtration step or one more step with PEG 6000 at 3.5% to avoid the contamination with lipids.

• Additionally, the changes in the type of agitation, centrifugation and the skip of dialysis make the method more accessible for all the laboratories.

• In summary, these modifications serve to enhance the purity, reduce the time for IgY extraction from egg yolk and make it more accessible for every basic research laboratory.

• Antibody
• Extraction
• Hen
• IgY
• PEG 6000

• Anti-microbial resistance
• chicken antibodies
• hospital acquired pneumonia (HAP)
• pneumonia

• Animals
• Immunoglobulins, Intravenous/therapeutic use
• Oxygen
• Pneumonia, Ventilator-Associated/drug therapy
• Pseudomonas aeruginosa
• Swine

• Region Örebro län
• VINNOVA

Since the discovery of antibodies by Emil Von Behring and Shibasaburo Kitasato during the 19th century, their potential for use as biotechnological reagents has been exploited in different fields, such as basic and applied research, diagnosis, and the treatment of multiple diseases. Antibodies are relatively easy to obtain from any species with an adaptive immune system, but birds are animals characterized by relatively easy care and maintenance. In addition, the antibodies they produce can be purified from the egg yolk, allowing a system for obtaining them without performing invasive practices, which favors the three “rs” of animal care in experimentation, i.e., replacing, reducing, and refining. In this work, we carry out a brief descriptive review of the most outstanding characteristics of so-called “IgY technology” and the use of IgY antibodies from birds for basic experimentation, diagnosis, and treatment of human beings and animals.

The novel severe acute respiratory syndrome (SARS) coronavirus, SARS-CoV-2, is responsible for the global COVID-19 pandemic. Effective interventions are urgently needed to mitigate the effects of COVID-19 and likely require multiple strategies. Egg-extracted antibody therapies are a low-cost and scalable strategy to protect at-risk individuals from SARS-CoV-2 infection. Commercial laying hens were hyperimmunized against the SARS-CoV-2 S1 protein using three different S1 recombinant proteins and three different doses. Sera and egg yolk were collected at three and six weeks after the second immunization for enzyme-linked immunosorbent assay and plaque-reduction neutralization assay to determine antigen-specific antibody titers and neutralizing antibody titers, respectively. In this study we demonstrate that hens hyperimmunized against the SARS-CoV-2 recombinant S1 and receptor binding domain (RBD) proteins produced neutralizing antibodies against SARS-CoV-2. We further demonstrate that antibody production was dependent on the dose and type of antigen administered. Our data suggests that antibodies purified from the egg yolk of hyperimmunized hens can be used as immunoprophylaxis in humans at risk of exposure to SARS-CoV-2.

• COVID-19
• SARS-CoV-2
• antibodies
• chicken
• egg
• neutralizing antibodies
• passive immunization

• COVID-19
• IgY
• SARS-CoV-2
• egg-yolk antibodies
• passive immunization
• receptor binding domain

• COVID-19
• IgY
• SARS-CoV-2
• chicken immunoglobulin
• clinical trial body
• immunoglobulin Y
• infectious diseases

• Animals
• Antibodies, Viral
• COVID-19/prevention & control
• Chickens
• Female
• Humans
• Immunoglobulins
• Rats
• SARS-CoV-2
• Spike Glycoprotein, Coronavirus

The urgent need for vaccines against Ebola virus (EBOV) was underscored by the large outbreak in West Africa (2014–2016). Since then, several promising vaccine candidates have been tested in pre-clinical and clinical studies. As a result, two vaccines were approved for human use in 2019/2020, of which one includes a heterologous adenovirus/Modified Vaccinia virus Ankara (MVA) prime-boost regimen. Here, we tested new vaccine candidates based on the recombinant MVA vector, encoding the EBOV nucleoprotein (MVA-EBOV-NP) or glycoprotein (MVA-EBOV-GP) for their efficacy after homologous prime-boost immunization in mice. Our aim was to investigate the role of each antigen in terms of efficacy and correlates of protection. Sera of mice vaccinated with MVA-EBOV-GP were virus-neutralizing and MVA-EBOV-NP immunization readily elicited interferon-γ-producing NP-specific CD8+ T cells. While mock-vaccinated mice succumbed to EBOV infection, all vaccinated mice survived and showed drastically decreased viral loads in sera and organs. In addition, MVA-EBOV-NP vaccinated mice became susceptible to lethal EBOV infection after depletion of CD8+ T cells prior to challenge. This study highlights the potential of MVA-based vaccines to elicit humoral immune responses as well as a strong and protective CD8+ T cell response and contributes to understanding the possible underlying mechanisms.

• Click reaction
• Copper-polydopamine nanoparticles
• DNAzyme
• Foodborne pathogen
• Microfluidic immunosensor
• Stir bar enrichment

• Helicobacter pylori
• IgY
• IgY−H. pylori
• alternative therapy
• egg yolk

• Animals
• Antibodies
• Egg Yolk
• Helicobacter pylori
• Immunoglobulins
• Urease

Egg yolk antibody (immunoglobulin Y, IgY), due to its unique features (e.g., cost-effectiveness for mass production), is emerging as a promising passive immune agent and alternative to antibiotics to combat infectious diseases, particularly in livestock. Oral administration of egg yolk IgY is the most common and convenient route that has been extensively investigated for controlling enteric pathogens. However, the in vivo stability of egg yolk IgY in the gastrointestinal (GI) tract, a critical issue for the success of this approach, still has not been clearly elucidated. Our recent study showed instability of orally administered egg yolk IgY in chicken GI tract, as demonstrated by both in vivo and ex vivo evidence. To better understand the magnitude and dynamics of instability of egg yolk IgY in vivo, in this study, we conducted comprehensive ex vivo analyses by spiking hyperimmune egg yolk IgY in fresh GI contents collected from five broilers at each sampling age (2, 4, or 6 weeks). The pH in gizzard slightly increased with age from 2.4 to 3.0, while the pH in the small intestine was around 5.8. ELISA analysis indicated that a short time of treatment (30 or 60 min) of IgY with the gizzard contents from the chickens at 2, 4, and 6 weeks of age greatly reduced specific IgY titer by over 8, 6, and 5 log₂ units, respectively, when compared with saline control. However, small intestine content only had a mild effect on egg yolk IgY, leading to 1 log₂ unit of reduction in IgY titer upon 30 min of treatment. Consistent with these findings, SDS-PAGE and immunoblotting analyses provided direct evidence demonstrating that egg yolk IgY could be drastically degraded to undetectable level in gizzard content upon as short as 5 min of treatment; however, the IgY was only slightly degraded in small intestine content. Immunoblotting also showed that treatment of IgY with HCl (pH 3.0) for 60 min did not affect its integrity at all, further supporting the enzymatic degradation of IgY in gizzard. Collectively, egg yolk IgY could be substantially degraded in chicken gizzard, highly warranting the development of effective approaches, such as encapsulation, for the controlled release and protection of orally administered egg yolk IgY in livestock.

• IgY stability
• chicken gizzard
• egg yolk antibody
• gastrointestinal digestion
• passive immunization

The term “IgY technology” was introduced in the literature in the mid 1990s to describe a procedure involving immunization of avian species, mainly laying hens and consequent isolation of the polyclonal IgYs from the “immune” egg yolk (thus avoiding bleeding and animal stress). IgYs have been applied to various fields of medicine and biotechnology. The present article will deal with specific aspects of IgY technology, focusing on the currently reported methods for developing, isolating, evaluating and storing polyclonal IgYs. Other topics such as current information on isolation protocols or evaluation of IgYs from different avian species are also discussed. Specific advantages of IgY technology (e.g., novel antibody specificities that may emerge via the avian immune system) will also be discussed. Recent in vitro applications of polyclonal egg yolk-derived IgYs to the field of disease diagnosis in human and veterinary medicine through in vitro immunodetection of target biomolecules will be presented. Moreover, ethical aspects associated with animal well-being as well as new promising approaches that are relevant to the original IgY technology (e.g., development of monoclonal IgYs and IgY-like antibodies through the phage display technique or in transgenic chickens) and future prospects in the area will also be mentioned.

• Animal welfare
• Polyclonal IgYs
• Egg yolk
• IgY technology
• Relevant-to-IgY-technology approaches
• In vitro immunodetection techniques

• Immunoglobulin Y (IgY); Microencapsulation; Chitooligosaccharide (COS); Response surface methodology (RSM); Controlled release; Simulated gastrointestinal digestion (SGID)

Besides being a common food component broadly consumed worldwide, egg yolk immunoglobulin Y (IgY) has essential therapeutic potentials. In fact, in a time of ever-increasing risk of antibiotic resistance, it is crucial to find new ways to battle infection, and oral administration of preformed specific antibodies represents one of the most attractive approaches against infection. Infectious diseases of bacterial and viral origin in humans and animals can be controlled and passively cured by orally applied IgYs isolated from chicken egg yolks. Despite multiple obvious advantages of oral administration of IgY, harvesting IgY from egg yolk in a pure form is a challenging task.

In this study, we developed a fast, simple, cost-effective, and efficient protocol for IgY isolation from chicken egg yolks. First, egg yolk was collected and diluted with 5 volumes of cold distilled water, homogenized, pH adjusted, and centrifuged. Next, the supernatant was collected, to which caprylic acid at concentration of 2% v/v was added, followed by pH adjustment to pH 5.0, centrifugation at 4°C, and collection of the resulting supernatant. This step was repeated twice, with adding 2% v/v of caprylic acid each time. The final supernatant was concentrated using ultrafiltration, and the IgY purity and activities were checked by SDS-PAGE, western blotting, and ELISA. The sequential (2, 2, 2%) addition of caprylic acid yielded IgY with a purity of 63.5, 90.6, and 95.8%, respectively, and reached 97.9% after ultrafiltration at pH 9.0. The IgY activity increased exponentially to reach 99% after the ultrafiltration step.

The proposed caprylic-acid-based protocol of IgY purification from the yolk of chicken eggs seems to be simple, fast, direct, and very cheap. This indicates that this protocol has great potential for scale-up processing.

• IgY
• benefits
• chicken
• purification
• structure

This new decade has started with a global pandemic of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), precipitating a worldwide health crisis and economic downturn. Scientists and clinicians have been racing against time to find therapies for COVID-19. Repurposing approved drugs, developing vaccines and employing passive immunization are three major therapeutic approaches to fighting COVID-19. Chicken immunoglobulin Y (IgY) has the potential to be used as neutralizing antibody against respiratory infections, and its advantages include high avidity, low risk of adverse immune responses, and easy local delivery by intranasal administration. In this study, we raised antibody against the spike (S) protein of SARS-CoV-2 in chickens and extracted IgY (called IgY-S) from egg yolk. IgY-S exhibited high immunoreactivity against SARS-CoV-2 S, and by epitope mapping, we found five linear epitopes of IgY-S in SARS-CoV-2 S, two of which are cross-reactive with SARS-CoV S. Notably, epitope SIIAYTMSL, one of the identified epitopes, partially overlaps the S1/S2 cleavage region in SARS-CoV-2 S and is located on the surface of S trimer in 3D structure, close to the S1/S2 cleavage site. Thus, antibody binding at this location could physically block the access of proteolytic enzymes to S1/S2 cleavage site and thereby impede S1/S2 proteolytic cleavage, which is crucial to subsequent virus-cell membrane fusion and viral cell entry. Therefore, the feasibility of using IgY-S or epitope SIIAYTMS-specific IgY as neutralizing antibody for preventing or treating SARS-CoV-2 infection is worth exploring.

Periodontitis is a bacterial biofilm-induced oral disease, mostly caused by Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) and Porphyromonas gingivalis (P. gingivalis). Oral administration of chicken egg yolk antibody (IgY) is a promising nutritional strategy to control pathogen infections. The objective of this study was to produce an A. actinomycetemcomitans- and P. gingivalis-specific IgY and evaluate its effects on bacterial agglutination and biofilm formation. Thirty laying hens were immunized with a complex of lysate containing typical molecular weights of membrane proteins of A. actinomycetemcomitans and P. gingivalis. IgY was isolated by polyethylene glycol 6000 and ammonium sulfate and purified by dialysis. The results of enzyme-linked immunosorbent assay showed that the obtained IgY were specific to both A. actinomycetemcomitans and P. gingivalis. In addition, immunoelectron microscopy scanning and crystal violet staining showed that the IgY could bind to cell wall of the pathogens and efficiently accelerate agglutination and inhibit biofilm formation. Furthermore, the activity of the IgY remained stable at different temperature, pH, and storage period. This is the first report that a novel two-in-one IgY was produced to modulate the agglutination and biofilm formation of A. actinomycetemcomitans and P. gingivalis, suggesting the potential of IgY to control periodontitis caused by oral pathogens.

• Agglutination
• Aggregatibacter actinomycetemcomitans
• Biofilm
• Egg yolk antibody
• Periodontitis
• Porphyromonas gingivalis

• Anti-protein A antibody
• Chicken IgY
• Gold nanoparticles
• Immunosensor
• Staphylococcus aureus

• anti-infective agents
• antibodies
• epitope
• immune therapy
• peptide
• vaccine

• APOGEO INTERREGMAC Call-European Funds for Regional Development-FEDER
• Immunowine TF INNOVA 2016-21-MEDI & FDCAN Funds by Cabildo de Tenerife

• Hedgehog
• RND transporter
• cation gradient
• cell signaling
• cholesterol transport

The world is facing one of the major outbreaks of viral infection of the modern history, however, as vaccine development workflow is still tedious and can not control the infection spreading, researchers are turning to passive immunization as a good and quick alternative to treat and contain the spreading. Within passive immunization domain, raising specific immunoglobulin (Ig)Y against acute respiratory tract infection has been developing for more than 20 years. Far from being an obsolete chapter we will revise the IgY-technology as a new frontier for research and clinic. A wide range of IgY applications has been effectively confirmed in both human and animal health. The molecular particularities of IgY give them functional advantages recommending them as good candidates in this endeavor. Obtaining specific IgY is sustained by reliable and nature friendly methodology as an alternative for mammalian antibodies. The aria of application is continuously enlarging from bacterial and viral infections to tumor biology. Specific anti-viral IgY were previously tested in several designs, thus its worth pointing out that in the actual COVID-19 pandemic context, respiratory infections need an enlarged arsenal of therapeutic approaches and clearly the roles of IgY should be exploited in depth.

• COVID-19
• IgY
• acute respiratory infection
• immunoglobulin

• Immunoglobulin Y (IgY)
• fluorescein isothiocyanate (FITC) labeling
• horseradish peroxidase (HRP) labeling
• mouse IgG
• secondary antibody

• IgY antibody
• Salmonella enterica subsp. enterica serovar Infantis
• antibacterial property
• broiler chicken
• capsulation

• Animal Feed/analysis
• Animals
• Antibodies, Bacterial/immunology
• Chickens
• Diet/veterinary
• Dietary Supplements/analysis
• Egg Yolk/physiology
• Female
• Immunoglobulins/immunology
• Intestines/microbiology
• Male
• Poultry Diseases/microbiology
• Poultry Diseases/prevention & control
• Random Allocation
• Salmonella Infections, Animal/microbiology
• Salmonella Infections, Animal/prevention & control
• Salmonella enterica/drug effects
• Salmonella enterica/physiology
• Serogroup

Gallibacterium anatis is a Gram-negative opportunistic avian pathogen representing an emerging threat to poultry meat and egg production worldwide. To date, no vaccine able to effectively prevent the morbidity associated with G. anatis infections has been developed yet. Our group previously reported that inoculation of different combinations of G. anatis outer membrane vesicles (OMVs), FlfA and GtxA-N proteins is effective in preventing lesions caused by G. anatis infections in layer chickens. Here we report the testing of the efficacy as vaccine prototypes of G. anatis OMVs isolated by hydrostatic filtration, a simple technique that allows the cost-effective isolation of high yields of OMVs. Layer chickens were immunized with OMVs alone or in combination with FlfA and/or GtxA-N proteins. Subsequent challenge with a heterologous G. anatis strain showed that immunization with OMVs alone could significantly reduce the lesions following a G. anatis infection. A second study was carried out to characterize the dose-response (0.25, 2.5 and 25 µg) relationship of G. anatis OMVs as immunogens, showing that 2.5 μg of OMVs represent the optimal dose to elicit protection in the immunized animals after a similar challenge. Additionally, administration of ≥2.5 μg of G. anatis OMVs induced specific IgY titers and possibly vertical transfer of immunity.

• Gallibacterium anatis
• hydrostatic filtration
• outer membrane vesicles
• vaccination

• ELISA methods
• Immunoglobulin Y
• alkaline phosphatase
• diagnosis
• influenza A

• Birds
• Diagnostics
• Immunoglobulin Y
• Immunotherapy
• Prophylaxis

• Animals
• Bioterrorism
• Egg Yolk/immunology
• Food Preservation
• Humans
• Immunoglobulins/chemistry
• Immunoglobulins/immunology
• Immunoglobulins/therapeutic use
• Infections/diagnosis
• Infections/immunology
• Neoplasms/diagnosis
• Neoplasms/immunology

• IgY
• fumonisin B
• lateral flow immunoassay
• rapid methods

• Animals
• Fumonisins/analysis
• Gold
• Humans
• Immunoassay
• Immunoglobulins
• Metal Nanoparticles
• Zea mays/chemistry
• Zea mays/microbiology

• IgY
• fumonisin B
• lateral flow immunoassay
• rapid methods

• Animals
• Fumonisins/analysis
• Gold
• Humans
• Immunoassay
• Immunoglobulins
• Metal Nanoparticles
• Zea mays/chemistry
• Zea mays/microbiology

• Egg yolk
• HRP
• Human serum
• IgY
• Polyclonal antibody

• Human norovirus
• IgY
• vesicular stomatitis virus (VSV)-based vaccine

• Animals
• Antibodies, Viral/blood
• Blood Group Antigens
• Caliciviridae Infections/immunology
• Capsid Proteins
• Chickens/immunology
• Egg Yolk/immunology
• Female
• Gastroenteritis/virology
• Gene Expression Regulation, Viral
• Humans
• Immunization
• Immunization, Passive
• Immunoglobulins/immunology
• Kinetics
• Norovirus/immunology
• Vaccination
• Vesicular Stomatitis/virology
• Viral Structural Proteins
• Viral Vaccines/immunology

• R01 AI123661 NIAID NIH HHS
• AI12366 NIH HHS

P. aeruginosa is a pathogen frequently resistant to antibiotics and a common cause of ventilator-associated pneumonia (VAP). Non-antibiotic strategies to prevent or treat VAP are therefore of major interest. Specific polyclonal avian IgY antibodies have previously been shown to be effective against pneumonia caused by P. aeruginosa in rodents and against P. aeruginosa airway colonization in patients.

To study the effect of specific polyclonal anti-P. aeruginosa IgY antibodies (Pa-IgY) on colonization of the airways in a porcine model.

The pigs were anesthetized, mechanically ventilated, and subject to invasive hemodynamic monitoring and allocated to either receive 10⁹ CFU nebulized P. aeruginosa (control, n = 6) or 10⁹ CFU nebulized P. aeruginosa + 200 mg Pa-IgY antibodies (intervention, n = 6). Physiological measurement, blood samples, and tracheal cultures were then secured regularly for 27 h, after which the pigs were sacrificed and lung biopsies were cultured.

After nebulization, tracheal growth of P. aeruginosa increased in both groups during the experiment, but with lower growth in the Pa-IgY-treated group during the experiment (p = 0.02). Tracheal growth was 4.6 × 10³ (9.1 × 10²–3.1 × 10⁴) vs. 4.8 × 10⁴ (7.5 × 10³–1.4 × 10⁵) CFU/mL in the intervention group vs. the control group at 1 h and 5.0 × 10⁰ (0.0 × 10⁰–3.8 × 10²) vs. 3.3 × 10⁴ (8.0 × 10³–1.4 × 10⁵) CFU/mL at 12 h in the same groups. During this time, growth in the intervention vs. control group was one to two orders of ten lower. After 12 h, the treatment effect disappeared and bacterial growth increased in both groups. The intervention group had lower body temperature and cardiac index and higher static compliance compared to the control group.

In this porcine model, Pa-IgY antibodies lessen bacterial colonization of the airways.

The online version of this article (10.1186/s40635-019-0246-1) contains supplementary material, which is available to authorized users.

• HAP
• Nosocomial
• Pneumonia
• VAP

• Chinese sacbrood virus
• IgY against CSBV
• antibody titer
• controlling of CSBV infection
• egg yolk antibodies
• passive immunotherapy
• preparation of IgY

• Fluorescence
• IgG
• IgY
• Immunomagnetic beads
• Quantum dots

• National Natural Science Foundation of China
• Graduate Innovation Fund of Jilin University

Adenosine deaminase (ADA) is currently used as a diagnostic marker for tuberculous pleuritis. Although ADA has been suggested as a potential marker for several types of cancer, the importance of each of ADA isoforms as well as their levels and enzymatic activities in tumors need to be further investigated. Herein we developed avian immunoglobulin Y highly specific to human ADA via hens immunization with calf adenosine deaminase. The obtained antibodies were used for the development of a sensitive double-egg yolk immunoglobulin (IgY) sandwich ELISA assay with an ADA detection limit of 0.5 ng/ml and a linearity range of up to 10 ng/ml. Specific, affinity-purified IgYs were able to recognize human recombinant ADA and ADA present in human cancer cell lines. In addition, antigen-specific IgY antibodies were able to inhibit catalytic activity of calf ADA with an IC₅₀ value of 47.48 nM. We showed that generated IgY antibodies may be useful for ADA detection, thus acting as a diagnostic agent in immunoenzymatic assays.

• Adenosine deaminase (ADA)
• Affinity purification
• Anti-adenosine deaminase antibody
• ELISA
• Egg yolk immunoglobulin (IgY)
• Enzyme inhibition