IgY technology: extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation.
IgY technology: extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation.
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DOI：
10.3791/3084
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发表时间：
2011-05-01
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影响因子：
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通讯作者：
Schade, Rudiger
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作者：
Pauly, Diana;Chacana, Pablo A;Schade, Rudiger
母鸡可通过免疫接种（胸肌，左、右，注射量0.5 ~ 1.0 ml）或通过Gene-Gun质粒免疫。根据抗原的免疫原性，只需1次或3 - 4次增强免疫即可获得高抗体滴度（高达1:10万- 1:10万）。正常情况下，母鸡连续下蛋约72周，之后产蛋量下降。本方案描述了用沉淀法（PEG）从蛋黄中提取总卵黄IgY的方法。Polson et al. 1980)。该方法包括两个重要步骤。第一步是去除脂质，第二步是从第一步的上清液中沉淀总卵磷脂。在缓冲液（通常是PBS）透析后，卵磷脂提取物可在-20°C下保存一年以上。提取物的纯度约为80%，每个鸡蛋的卵黄素含量在40-80毫克之间，这取决于蛋鸡的年龄。卵黄素总含量随着母鸡年龄的增长而增加，从大约40毫克/个鸡蛋增加到80毫克/个鸡蛋（涉及PEG析出）。一只母鸡每年的产蛋量约为325个蛋。这意味着基于平均IgY含量为60毫克/个鸡蛋，总IgY的总潜在收获量为20克/年（见表1）。
Hens can be immunized by means of i.m. vaccination (Musculus pectoralis, left and right, injection volume 0.5-1.0 ml) or by means of Gene-Gun plasmid-immunization. Dependent on the immunogenicity of the antigen, high antibody-titres (up to 1:100,000 - 1:1,000,000) can be achieved after only one or 3 - 4 boost immunizations. Normally, a hen lays eggs continuously for about 72 weeks, thereafter the laying capacity decreases. This protocol describes the extraction of total IgY from egg yolk by means of a precipitation procedure (PEG. Polson et al. 1980). The method involves two important steps. The first one is the removal of lipids and the second is the precipitation of total IgY from the supernatant of step one. After dialysis against a buffer (normally PBS) the IgY-extract can be stored at -20°C for more than a year. The purity of the extract is around 80 %, the total IgY per egg varies from 40-80 mg, dependent on the age of the laying hen. The total IgY content increases with the age of the hen from around 40 mg/egg up to 80 mg/egg (concerning PEG precipitation). The laying capacity of a hen per year is around 325 eggs. That means a total potential harvest of 20 g total IgY/year based on a mean IgY content of 60 mg total IgY/egg (see Table 1).