Department of Medical Sciences, Uppsala University, Uppsala University Hospital, Uppsala, Sweden.
NABAS AS, Ås, Norway.
Front Immunol. 2022 Oct 11;13:1016781. doi: 10.3389/fimmu.2022.1016781. eCollection 2022.
Antibody-based assays are commonly used in clinical laboratories for analyzing plasma, serum and other samples for particular protein markers. Although such assays have been traditionally based on antibodies raised in mammals (e.g., mice, rabbits, goats), there are several advantages of using avian antibodies (IgY) raised in chickens, including production volumes, costs, and ethical/animal welfare considerations. A further disadvantage of using mammalian IgG in such assays is the potential for agglutination when exposed to rheumatoid factor (RF) in serum. However, when used in the free form the immune complexes formed with avian antibodies have been reported to have less ability than those formed with mammalian antibodies to cause the light scatter which are used for instrument measurement. In addition, when the amount of antigen exceeds the maximum precipitating point in relation to the amount of antibody, there is a rapid decline in the absorbance values of the immune complexes (antigen excess) when IgY is used. However, when avian antibodies are conjugated to a substrate and used in particle enhanced turbidimetric assays (PETIA), these problems are avoided. Here we investigated three clinical assays using chicken antibodies, one using free (unbound) IgY and two with IgY-based PETIA. The IgY PETIA demonstrated a strong scatter response, even at high antigen concentrations in contrast to the steep decline seen with free IgY antibodies. IgY PETIA reagents can provide test results with low coefficient of variation (<1% for duplicate samples). We also investigated the effect of RF on agglutination of mammalian antibodies (IgG from mouse, rabbit, sheep, and human) and chicken antibodies. Whereas agglutination was observed with all the mammalian antibodies in the presence of RF, this was not observed at all with chicken IgY. Our results support the growing body of evidence that chicken egg yolks can thus be a valuable source of antibodies for use in PETIA in clinical laboratories.
基于抗体的检测方法常用于临床实验室，用于分析血浆、血清和其他样本中的特定蛋白质标志物。尽管此类检测方法传统上基于在哺乳动物（例如，小鼠、兔子、山羊）中产生的抗体，但使用在鸡中产生的禽类抗体（IgY）具有几个优点，包括生产体积、成本以及伦理/动物福利方面的考虑。在这些检测中使用哺乳动物 IgG 的另一个缺点是在血清中暴露于类风湿因子 (RF) 时可能会发生凝集。然而，当以游离形式使用时，据报道与用哺乳动物抗体形成的免疫复合物相比，与禽类抗体形成的免疫复合物形成的光散射的能力更小，这些光散射用于仪器测量。此外，当抗原的量超过与抗体的最大沉淀点相关的量时，当使用 IgY 时，免疫复合物的吸光度值（抗原过量）会迅速下降。然而，当禽类抗体与基质偶联并用于颗粒增强浊度测定法 (PETIA) 时，这些问题就会得到避免。在这里，我们研究了三种使用鸡抗体的临床检测方法，一种使用游离（未结合）IgY，两种使用基于 IgY 的 PETIA。与游离 IgY 抗体观察到的急剧下降相比，IgY PETIA 显示出很强的散射反应，即使在高抗原浓度下也是如此。IgY PETIA 试剂可以提供具有低变异系数（<1%的重复样本）的测试结果。我们还研究了 RF 对哺乳动物抗体（来自小鼠、兔子、绵羊和人类的 IgG）和鸡抗体的凝集作用的影响。虽然在存在 RF 的情况下观察到所有哺乳动物抗体的凝集，但在鸡 IgY 中完全没有观察到这种情况。我们的结果支持越来越多的证据表明，鸡卵黄可以成为用于临床实验室 PETIA 的抗体的有价值来源。
