Preparation of Immunoglobulin Y from Egg Yolk Using Ammonium Sulfate Precipitation and Ion Exchange Chromatography

Ko, Kyung Yuk; Ahn, Dong U.

doi:10.1093/ps/86.2.400

Cited by 83 publications

(58 citation statements)

References 37 publications

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“…Solution was precipitated one night on 4° C and was centrifuged with 10,000 rpm for an hour. 9 Precipitate was taken for re-suspension using PBS then it was purified and analyzed. Immunoglobulin Y precipitation was covered with specific plastic for sonication, then added 0.5M PBS and stirred using magnetic stirrer for 24 hours in 4° C. Characterization was done by reacted antibody from purification with antigen membrane using ELISA method and Western Bloth.…”

Section: Methodsmentioning

confidence: 99%

Production and Characterization of Immunoglobuline Yolk as anti antigen membrane Toxoplasma gondii

Puspitasari

Praptiwi

Suwanti‬

2016

IJTID

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Section: Methodsmentioning

confidence: 99%

Production and Characterization of Immunoglobuline Yolk as anti antigen membrane Toxoplasma gondii

Puspitasari

Praptiwi

Suwanti‬

2016

IJTID

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“…IgY was purified from lyophilized egg yolk powder essentially as described by Ko & Ahn (2007). Briefly, 2 g lyophilized egg yolk powder was dissolved in 150 ml ice-cold distilled water.…”

Section: Methodsmentioning

confidence: 99%

“…FliD-specific IgY was purified from lyophilized egg yolks using the two-step charcoal de-lipidation ammonium sulfate precipitation process previously reported by Ko & Ahn (2007). SDS-PAGE analysis (Fig.…”

Section: Fig 1 Is An Image Of An Sds-page Gel On Which Wementioning

confidence: 99%

Therapeutic potential of egg yolk antibodies for treating Clostridium difficile infection

Mulvey

Dingle

Fang³

et al. 2011

Journal of Medical Microbiology

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Herein we present evidence for the therapeutic potential of colonization factor (CF)-specific egg yolk antibodies (IgY) for potentially treating acute and recurring Clostridium difficile infection (CDI) in humans. The study involved cloning, expressing as 6¾His-tagged proteins in Escherichia coli, and Ni-affinity purifying three previously identified CFs (FliC, FliD and Cwp84) from C. difficile. The recombinant CF antigens were then used to immunize Leghorn chickens and CF-specific IgY antibodies were prepared from their eggs. The specificity and titre of the resulting C. difficile CF-specific IgY antibodies were assessed by ELISA and Western immunoblotting techniques. The antibodies were also screened for their ability to inhibit C. difficile adherence to human colon-derived T84 cells, and, based on these findings, one of them (FliD-specific IgY) was evaluated for its potential to prevent C. difficile-mediated morbidity and mortality in Syrian hamsters. The results revealed that purified FliD-specific IgY significantly protected hamsters from C. difficile strain 630 infection relative to control animals treated with carbonate buffer alone or IgY produced from unimmunized chicken eggs. The results suggest that egg yolk preparations obtained from chickens immunized with recombinant C. difficile CFs may represent another safe and cost-effective treatment option in humans suffering from acute or recurring CDI. INTRODUCTIONConventional antibiotics alter both the numbers and complexity of the normal gastrointestinal (GI) microflora, thereby reducing the ability of these protective organisms to competitively exclude opportunists from colonizing the GI tract. This disruption may render hospitalized subjects susceptible to colonization by C. difficile (Job & Jacobs, 1997). Once established in the colon, C. difficile expresses two very large exotoxins, toxin A (TcdA) and toxin B (TcdB), which are primarily responsible for the clinical symptoms of C. difficile infection (CDI) (Voth & Ballard, 2005). These range from diarrhoea to pseudomembranous colitis, toxic megacolon and multi-system organ failure, which may be fatal.The accepted approach to treating CDI is to replace the antibiotic responsible for initiating the condition with one, typically metronidazole, to which C. difficile is sensitive. In about 80 % of cases, this approach leads to complete recovery without further complications. In the remaining cases, however, recurring CDI (rCDI) can occur, due, presumably, to a failure of the normal GI microflora to become re-established after the metronidazole is discontinued. This leaves the colon of these individuals susceptible to C. difficile relapse or reinfection. Multiple recurrences are not unusual, resulting in a significant reduction in the quality of life in these individuals.In response to the increasing risk of CDI to human health, much research has focused on new and improved treatment regimens, particularly in subjects suffering from rCDI. These studies have concentrated on novel toxin neutralizing agents, an...

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“…The bound IgY was eluted with 200 mM NaCl in Tris 10 mM. The eluate was collected, dialysed and stored until used (Ko and Ahn 2007).…”

Section: Methodsmentioning

confidence: 99%

Development of Rapid Agglutination Test to Detect Chicken Marek Antibody

Wibawan¹,

Halimah²,

Djannatun³

et al. 2009

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To our knowlege, there is no rapid agglutination test to detect antibodies to viruses which might be due to the small dimension of viral particles. Through complex formation of Staphylococcus aureus bearing protein A-rabbit IgG-anti IgY-IgY anti Marek virus, agglutination of antibodies to viruses can be achieved and visualized. To design the prototype of the test, the bacterial cells of Staphylococcus aureus were coupled to a complex compound consisting of IgG-IgY-Marek antigen. This protocol was able to detect clearly the presence of Marek antibody in chicken sera, showing the rapid, clear and distinct agglutination reaction on the glass objects. No agglutination reaction was observed in the reaction of specified pathogen free chicken sera with this prototype showing the specificity of the test. This finding demonstrates a novel rapid agglutination which can be used for the detection of antibodies to various agents.

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Preparation of Immunoglobulin Y from Egg Yolk Using Ammonium Sulfate Precipitation and Ion Exchange Chromatography

Cited by 83 publications

References 37 publications

Production and Characterization of Immunoglobuline Yolk as anti antigen membrane Toxoplasma gondii

Production and Characterization of Immunoglobuline Yolk as anti antigen membrane Toxoplasma gondii

Therapeutic potential of egg yolk antibodies for treating Clostridium difficile infection

Development of Rapid Agglutination Test to Detect Chicken Marek Antibody

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