The purification of IgY from chicken egg yolk by preparative electrophoresis

Gee, Sarah; Bate, Irene M; Thomas, Theresa M.; Rylatt, Dennis B.

doi:10.1016/s1046-5928(03)00076-7

Cited by 19 publications

(12 citation statements)

References 19 publications

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“…The isoelectric point of IgY was measured by IEF. The pI of IgY was pH 6.2-6.5, which was close to that of the literature value (6.5-7.5) (Gee et al, 2003).…”

Section: Electrophoresissupporting

confidence: 89%

“…Although the importance of chicken egg as an economical and convenient source of polyclonal antibodies has been well recognised, the widespread use is often considered impractical partly due to the difficulties in isolation of IgY from yolk (Devi et al, 2002;Kuroki, 1999). The conventional techniques such as precipitation (Akita & Nakai, 1993), electrophoresis (Gee, Bate, Thomas, & Rylatt, 2003), and chromatography (Hansen, Scoble, Hanson, & Hoogenraad, 1998;Tini, Jewell, Camenisch, Chilov, & Gassmann, 2002;Verdoliva, Basile, & Fassina, 2000) have been used in the IgY separation and purification process. However, these conventional techniques are often time consuming and low yielding (Verdoliva et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

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A new membrane based process to isolate immunoglobulin from chicken egg yolk

Liu

Yang

et al. 2010

Food Chemistry

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a b s t r a c tAlthough the importance of eggs as a source of specific antibodies has been well recognised, the generation of egg yolk immunoglobulins (IgY) is rarely chosen due to the peculiar composition of egg yolk and the lack of specific affinity ligands. In this work, we report a novel membrane based two-stage ultrafiltration process to isolate IgY from egg yolk. The effects of solution pH, ionic strength, stirring speed and permeate flux on the transmission of proteins were quantified using the pulsed sample injection technique and parameter scanning ultrafiltration. Under optimised conditions, the purity of immunoglobulin obtained was greater than 93% after the two-stage ultrafiltration process and the recovery of immunoglobulin from the feedstock was close to 87%. The resulting immunoglobulin product was then analysed by Isoelectric Focusing (IEF), SDS-PAGE and Circular Dichroism (CD), to confirm its isoelectric point, molecular weight and molecular secondary structure.

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“…The isoelectric point of IgY was measured by IEF. The pI of IgY was pH 6.2-6.5, which was close to that of the literature value (6.5-7.5) (Gee et al, 2003).…”

Section: Electrophoresissupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

A new membrane based process to isolate immunoglobulin from chicken egg yolk

Liu

Yang

et al. 2010

Food Chemistry

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“…This result indicates that extraction method also affect the protein concentration by the salting precipitation. Following, there are many methods to purify the IgY from yolk (Schwarzkopf & Thiele, 1996;Gee et al, 2003). Firstly, simple water dilution method is used to remove lipoproteins from WSF for the IgY purification (Jensenius et al, 1981;Kwan et al, 1991;Akita & Nakai, 1992;1993).…”

Section: Resultsmentioning

confidence: 99%

Extraction and Purification of Anti-Helicobacter pylori IgY

Siriya¹,

Chu²,

Chen³

et al. 2013

JAS

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The aim of this study was to establish an efficient procedure to produce anti-H. pylori urease IgY from the eggs of the laying hens. Four White Leghorn hens (52 weeks old) were immunized intramuscularly with the purified urease of H. pylori with aluminum hydroxide gel adjuvant. To increase the specificity and antibody titer, three boosters were given at 2-week intervals following the first injection. After the final immunization, the eggs were collected daily and stored at 4°C. The yield of yolk proteins were 47.52%, 41.07% and 51.35% for the methods of dextran sulphate, isopropanol and water-soluble fraction (WSF), respectively. Further, IgY was purified successively by ammonia sulfate, affinity chromatography and ion exchange chromatography for each crude protein. Ammonia sulfate salting produced the IgY with a purity of 37-53% and the titer of 22.6-29.5 unit/mg. The purity, relative purification folds and titer characteristics of the purified IgY were 85-90%, 19.63-34.61 and 223.8-273.4 unit/mg protein for affinity chromatography, and 89-92%, 25.33-47.14 and 288.8-372.4 unit/mg protein for ion exchange chromatography, respectively. This anti-Helicobacter pylori IgY was specific against H. pylori usease. Here, we established a simple and inexpensive method to obtain a yield of 90% purity of the anti-H. pylori IgY in combinations of the WSF method, ammonia sulfate precipitation, and affinity chromatography.

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“…In the literature, several methods are proposed for the extraction and purification of IgY [18][19][20]; we followed the extraction proposed in [20] that employs 0.2 M acetate buffer at pH 5. Briefly, after mechanical separation of yolk and egg white, a known amount of yolk is dissolved in the buffer to obtain a starting concentration of 0.5 g/mL of egg yolk, then the solution is mixed with vortex and left for two hours in a conical centrifuge tube, to let the heavy and not-soluble fraction deposit.…”

Section: Samples Preparationmentioning

confidence: 99%

Electrochemical Immunosensor for Detection of IgY in Food and Food Supplements

et al. 2017

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Immunoglobulin Y is a water-soluble protein present in high concentration in hen serum and egg yolk. IgY has applications in many fields, e.g., from food stuff to the mass production of antibodies. In this work, we have implemented an electrochemical immunosensor for IgY based on templated nanoelectrodes ensembles. IgY is captured by the templating polycarbonate and reacted with anti-IgY labeled with horseradish peroxidase. In the presence of H 2 O 2 and methylene blue as the redox mediator, an electrocatalytic current is generated which scales with IgY concentration in the sample. After optimizing the extracting procedure, the immunosensor was applied for analysis of fresh eggs and food integrators. The data obtained with the biosensor were validated by SDS-PAGE and Western blot measurements.

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The purification of IgY from chicken egg yolk by preparative electrophoresis

Cited by 19 publications

References 19 publications

A new membrane based process to isolate immunoglobulin from chicken egg yolk

A new membrane based process to isolate immunoglobulin from chicken egg yolk

Extraction and Purification of Anti-Helicobacter pylori IgY

Electrochemical Immunosensor for Detection of IgY in Food and Food Supplements

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