Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA)

Vaillant, Angel Justiz; Anderson, Norma McFarlane; Mohammed, Brian Wisdom Wayne

doi:10.4172/2155-9872.1000175

Cited by 8 publications

(14 citation statements)

References 11 publications

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“…The 35 ELISAs were highly reproducible and this is a confirmatory results for some of the ELISAs that have been reported previously as direct (SpL, SpA, SpLA), and sandwich ELISA as SpG-SpLA assay [9,20,25]. The reason why we report that these tests were highly reproducible is the fact that their coefficient of variation (both intra-assays and inter-assays) were within the normal limits except for few immunoglobulin samples.…”

Section: Resultssupporting

confidence: 88%

“…The chicken IgY did not bind to SpA. Some of these interactions have been previously cited [2,9,6,25].…”

Section: Resultsmentioning

confidence: 66%

“…This binding does not interfere with the antigen binding sites on the immunoglobulin receptors. These receptors have been called immunoglobulin-binding protein, IBP [9–14]. Protein A and G have been used as immunological tools in serological tests used in the immunodiagnosis of infectious diseases, such as Borrelia burgdorferi in zoo animals [14].…”

Section: Introductionmentioning

confidence: 99%

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Use of immunoglobulin-binding bacterial proteins in immunodetection

Vaillant

2020

Preprint

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One of the aim of this study was to make universal chimeric conjugates to react with both avian and mammalian immunoglobulins in enzyme-linked immunosorbent assays (ELISAs). The periodate method was used in the conjugation process of cross-linking horseradish peroxidase to immunoglobulin-binding proteins (IBP) including staphylococcal protein A (SpA), streptococcal protein G (SpG) and peptostreptococcal protein L (SpL). By mixing up these three conjugates another four hybrid protein conjugates were created including protein LA (SpLA), protein LG (SpLG), protein AG (SpAG) and protein LAG (PLAG). Thirty-five ELISAs were standardized by a probabilistic combination of these immunoreagents. By using a panel of mainly mammalian immunoglobulins their reproducibility was checked by the determination of coefficient of variations (CV) for each one of the IgG-IBP binding. The source of immunoglobulins was their purification by affinity chromatography using a commercially available kit (PURE-1A). The other aim was to immunize chicken with the peptide fragment 254-274 of gp120 to produce anti-HIV peptide hyper-immune egg. Cats and rats were fed these eggs for a determined period until they produced the anti-HIV peptide antibody, which was tested by an indirect SpLA-ELISA and dot blot analysis that corroborated the production of anti-HIV antibodies by the mammalian species including positive humans samples for HIV. We conclude that the single and hybrid immunoglobulin-binding protein were effective in their binding capacity to immunoglobulins from a variety of mammalian species. The potential use of this proteins is in the arena of immunodiagnosis and immunoglobulin detection. Dot blot analysis proves effective in the detection of HIV anti-gp120 antibodies in several animal species. These antibodies can be used as reagents in the development of immunodiagnostic tests or experimental vaccines.

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Section: Resultssupporting

confidence: 88%

“…The chicken IgY did not bind to SpA. Some of these interactions have been previously cited [2,9,6,25].…”

Section: Resultsmentioning

confidence: 66%

Section: Introductionmentioning

confidence: 99%

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Use of immunoglobulin-binding bacterial proteins in immunodetection

Vaillant

2020

Preprint

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“…In the case that is presented here, anti-Salmonella antibodies are detected in five species including a mammal (human) and four avian species more with only one standardized universal ELISA. The turtle IgY is used as a negative control as it does not bind to any of the immunoglobulin-binding proteins [21,24,25].…”

Section: Resultsmentioning

confidence: 99%

“…By chemical protein conjugation we engineered various hybrid bacterial Ig receptors with the capacity to bind to many immunoglobulins from mammalian and avian species. It includes SpLA-LG-peroxidase and SpLAG-anti-IgY-peroxidase produced by the periodate method [5]. These molecules display a very low background in immunoassays.…”

Section: Introductionmentioning

confidence: 99%

Universal enzyme-linked immunosorbent assays (ELISA) and utility in the detection of antibodies against Salmonella spp. in several animal species

Justiz-Vaillant¹,

Ferrer-Cosme²,

Curtello

2020

Preprint

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The aims of this research are confirming the feasibility of hybrid immunoglobulin-binding reagents, its used in ELISAs for IgG/IgY detection and detecting specific antibodies against an infectious microorganism (Salmonella spp) in various animal species using a universal diagnostic ELISA. Hybrid immunoglobulin-binding bacterial proteins (IBP) including recombinant protein LA, recombinant protein LG, and recombinant protein AG have been produced for improvement of their binding affinity to a much larger number of immunoglobulins. This hybrid bacterial protein thus represents a powerful tool for binding, detection, and purification of immunoglobulins and their fragments. However, SpLA-LG-peroxidase and SpLAG-anti-IgY-peroxidase were produced by the periodate method. They have shown to be effective reagents. Their binding affinity to immunoglobulins surpasses previous hybrid IgG-binding proteins reported, including the most known SpAG, SpLA and SpLG. The IgY fraction was isolated from the egg yolks of a variety of birds including species of chicken, bantam hen, guinea hen, quail, goose, duck, wild and domestic pigeon, parakeet, cattle egret, pheasant, and ostrich. The IgY fraction was isolated by the chloroform-polyethylene glycol (PEG) method. An ELISA for anti-Salmonella spp antibodies was employed with some modifications to determine the presence of antibodies in humans, laying hens, geese, quails, and pigeons. Salmonella are motile, flagellated rod-shaped zoonotic pathogens which may survive with or without oxygen. They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses. This pathogen is associated with several diseases, which may become fatal and negatively impact the health of individuals and various economies globally. The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.

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Preparation of horseradish peroxidase (HRP) conjugated to chicken anti-IgY v1

Justiz-Vaillant¹

2020

Preprint

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Avian IgY is structurally and genetically different from its mammalian counterpart IgG. This reagent can be used in ELISA, Western blotting and Dot blot to detect specific chicken IgY to infectious agents or immune system proteins [1]. IgY can be isolated by the Polson Method [2] or the water dilution method [3]. Chicken IgY can crossed-react with IgY from diverse bird species [4]. Similarly avian IgY can be isolated from their egg yolk using the same methodology with modifications that was developed for the chicken IgY isolation [5].

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Immunoglobulin-binding Bacterial Proteins (IBP) Conjugates and their Reactivity with Immunoglobulin in Enzyme-Linked Immunosorbent Assays (ELISA)

Cited by 8 publications

References 11 publications

Use of immunoglobulin-binding bacterial proteins in immunodetection

Use of immunoglobulin-binding bacterial proteins in immunodetection

Universal enzyme-linked immunosorbent assays (ELISA) and utility in the detection of antibodies against Salmonella spp. in several animal species

Preparation of horseradish peroxidase (HRP) conjugated to chicken anti-IgY v1

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