González R, Duffort O, Calabozo B, Barber D, Carreira J, Polo F
Research and Development Department, ALK-Abelló SA, Madrid, Spain.
Allergy. 2000 Jan;55(1):59-64. doi: 10.1034/j.1398-9995.2000.00366.x.
The demonstration that some asthma epidemics have been caused by allergens of soybean-hull dust prompted us to develop a two-site ELISA, suitable for the quantification of the major allergen (Gly m 1), to be used for the prevention of new episodes.
BALB/c mice were injected with Gly m 1 purified from soybean hulls. After fusion and screening, 10 monoclonal antibodies (mAbs) were obtained that were shown to be specific for Gly m 1 Two of them (6G1 as the capture antibody; 1G10 as the tracer) were selected to develop a quantitative two-site ELISA for the indoor and outdoor determination of Gly m 1.
The two-site ELISA developed is very sensitive, with a detection limit of less than 0.2 ng/ml and a practical working range of 0.4-10 ng/ml. The assay is also highly reproducible with an intra-assay coefficient of variation of 3.5% and an interassay coefficient of variation of 12.5%. The method was applied to measure the concentration of Gly m 1 in air-sampler filters and in house-dust samples. Our results illustrate that there is a good correlation between the content of Gly m 1 in a number of samples and the allergenic activity as measured by ELISA inhibition.
A specific and sensitive method is presented that can be used for the quantification of Gly m 1. The application of this method may allow the establishment of risk limits for soybean dust, and thus may contribute to the control of environmental contamination and to the prevention of new asthma epidemics.
一些哮喘流行是由大豆壳尘过敏原引起的，这一发现促使我们开发一种双位点酶联免疫吸附测定法（ELISA），用于定量主要过敏原（Gly m 1），以预防新的发病情况。
将从大豆壳中纯化的Gly m 1注射到BALB/c小鼠体内。经过融合和筛选，获得了10种单克隆抗体（mAb），它们被证明对Gly m 1具有特异性。其中两种（6G1作为捕获抗体；1G10作为示踪剂）被选来开发一种用于室内和室外定量测定Gly m 1的双位点ELISA。
所开发的双位点ELISA非常灵敏，检测限小于0.2 ng/ml，实际工作范围为0.4 - 10 ng/ml。该测定法的重复性也很高，批内变异系数为3.5%，批间变异系数为12.5%。该方法被应用于测量空气采样器过滤器和室内灰尘样本中Gly m 1的浓度。我们的结果表明，许多样本中Gly m 1的含量与通过ELISA抑制法测量的过敏活性之间存在良好的相关性。
提出了一种可用于定量Gly m 1的特异性和灵敏的方法。该方法的应用可能有助于确定大豆粉尘的风险限值，从而有助于控制环境污染和预防新的哮喘流行。
