Schranner I, Lösch U
Poult Sci. 1986 Feb;65(2):360-8. doi: 10.3382/ps.0650360.
Naturally existing variants of Immunoglobulin M (IgM) and Immunoglobulin A (IgA) in the serum of immunologically competent, as well as immunologically defective UM-B19 chickens can be detected without time consuming purification. The different molecular forms of IgM and IgA were separated by gel filtration. Pentameric IgM and dimeric IgA were well separated from the 7S-peak that contained monomeric IgM and IgA. A method for immunological identification of the protein components is described. Pore gradient gel electrophoresis was combined with antigen-antibody crossed electrophoresis in horizontal agarose slabs. The principle of the method is that IgM and IgA in their different molecular forms in the mixtures to be assayed after gel filtration are first separated with high resolution by sodium dodecylsulfate (SDS) polyacrylamide gel electrophoresis using a pore gradient of 3 to 15%. In the second step, a slab of polyacrylamide gel with the separated proteins in transferred onto agarose layers containing appropriate anti- mu- and anti-alpha antiserum and SDS in a concentration of .01%. The technique is easy to perform and gives reproducible results. In chicken serum, the monomeric state of IgM (184,000 d), along with the pentameric state (920,000 d) were identified. Serum IgA exists in dimeric (340,000 d) and in monomeric (170,000 d) states. No differences between the commercial Leghorn and dysgammaglobulinemic and normogammaglobulinemic lines were found.
在免疫功能正常以及免疫缺陷的UM - B19鸡血清中，无需耗时纯化就能检测到免疫球蛋白M（IgM）和免疫球蛋白A（IgA）的天然存在变体。通过凝胶过滤分离IgM和IgA的不同分子形式。五聚体IgM和二聚体IgA与含有单体IgM和IgA的7S峰得到了很好的分离。描述了一种蛋白质成分的免疫学鉴定方法。将孔径梯度凝胶电泳与水平琼脂糖平板中的抗原 - 抗体交叉电泳相结合。该方法的原理是，在凝胶过滤后的待检测混合物中，不同分子形式的IgM和IgA首先通过使用3%至15%孔径梯度的十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳进行高分辨率分离。第二步，将含有已分离蛋白质的聚丙烯酰胺凝胶平板转移到含有适当抗μ和抗α抗血清以及浓度为0.01% SDS的琼脂糖层上。该技术易于操作且结果可重复。在鸡血清中，鉴定出了IgM的单体状态（184,000 d）以及五聚体状态（920,000 d）。血清IgA以二聚体（340,000 d）和单体（170,000 d）状态存在。未发现商品来亨鸡与γ球蛋白血症和正常γ球蛋白血症品系之间存在差异。
