A novel monoclonal antibody screening method using the Luminex-100™ microsphere system
A novel monoclonal antibody screening method using the Luminex-100™ microsphere system
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DOI：
10.1016/s0022-1759(02)00168-0
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发表时间：
2002-09-15
影响因子：
2.2
通讯作者：
Peritt, D
中科院分区：
文献类型：
--
作者：
Seideman, J;Peritt, D
我们描述了使用 Luminex-100 微球开发的稳健且灵敏的测定系统（检测限 < 500 mug/ml 生物素-IL-6，K-d = 75 ng/ml），只要目标分子的受体的可溶形式可用，该系统就可以有效筛选中和抗体。例如，我们将重组人白细胞介素 6 可溶性受体与 Luminex 羧化微球偶联，并使用生物素标记的重组人白细胞介素 6 作为探针来评估结合竞争。使用结合不同IL-6表位的三种抗人IL-6单克隆抗体作为测试物品以证明筛选的严格性。我们的测定能够检测低至 10 ng/ml 的抗体浓度，而不受杂交瘤生长培养基或细胞上清液的干扰。这种检测形式具有节省时间的优点，使其成为中和单克隆抗体的高通量筛选 (HTS) 应用的理想选择。 (C) 2002 Elsevier Science B.V. 保留所有权利。
We describe the development of a robust and sensitive assay system (detection limit < 500 mug/ml biotin-IL-6, K-d = 75 ng/ ml), using Luminex-100 microspheres, that could effectively screen for neutralizing antibody whenever a soluble form of the receptor for a target molecule is available. As an example, we coupled a recombinant human interleukin-6 soluble receptor to a Luminex carboxylated microsphere and used a biotin-labeled recombinant human interleukin-6 as a probe to assess binding competition. Three anti-human IL-6 monoclonal antibodies that bind distinct IL-6 epitopes were used as test articles to evince the stringency of the screen. Our assay was able to detect antibody concentration as low as 10 ng/ml without interference from hybridoma growth medium or cell supernatant. The time-saving benefits of this assay format make it ideal for high-throughput screening (HTS) applications for neutralizing monoclonal antibodies. (C) 2002 Elsevier Science B.V. All rights reserved.