Brissac C, Nobrega A, Carneiro J, Stewart J
Unité d'Immunobiologie, Institut Pasteur, 25 rue du Dr Roux, 75015 Paris, France.
Int Immunol. 1999 Sep;11(9):1501-7. doi: 10.1093/intimm/11.9.1501.
This paper proposes a method for the quantitative characterization of repertoire diversity of an unknown mixture of antibodies on the basis of its reactivity profile in the quantitative immunoblot (QIB). The QIB is calibrated by measuring the reactivity profiles of supernatants of known 'diversity' (i.e. known numbers of B cell clones). We define a quantitative 'index of variability' (IV) which decreases regularly as the diversity increases and the profiles tend towards a common 'convergence profile'. The calibration procedure is consolidated by a mathematical model based on the Poisson distribution; this theoretical model accounts correctly for the observed convergence behavior. On the basis of this calibration curve, it is possible to estimate the diversity of an unknown antibody mixture from a measure of its IV. We conclude that the functional diversity of natural serum IgM in mice can be estimated at approximately 16,000 clones.
本文提出了一种基于未知抗体混合物在定量免疫印迹（QIB）中的反应谱对其库多样性进行定量表征的方法。通过测量已知“多样性”（即已知数量的B细胞克隆）的上清液的反应谱来校准QIB。我们定义了一个定量的“变异性指数”（IV），它随着多样性的增加而有规律地降低，并且反应谱趋向于一个共同的“收敛谱”。校准过程由基于泊松分布的数学模型巩固；这个理论模型正确地解释了观察到的收敛行为。基于这条校准曲线，通过测量未知抗体混合物的IV就可以估计其多样性。我们得出结论，小鼠天然血清IgM的功能多样性估计约为16,000个克隆。
