High-Throughput Screening of Single-Chain Antibodies Using Multiplexed Flow Cytometry

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• 作者：Joanne Ayriss ; Travis Woods ; Andrew Bradbury ; Peter Pavlik
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：March 2007
• 年：2007
• 卷：6
• 期：3
• 页码：1072 - 1082
• 全文大小：600K
• 年卷期：v.6,no.3(March 2007)
• ISSN：1535-3907

文摘

We have developed a screening method that has the potential to streamline the high-throughput analysisof affinity reagents for proteomic projects. By using multiplexed flow cytometry, we can simultaneouslydetermine the relative expression levels, the identification of nonspecific binding, and the discriminationof fine specificities to generate a complete functional profile for each clone. The quality and quantityof data, combined with significant reductions in analysis time and antigen consumption, provide notableadvantages over standard ELISA methods and yield much information in the primary screen which isusually only obtained in later screens. By combining high-throughput screening capabilities withmultiplex technology, we have redefined the parameters for the initial identification of affinity reagentsrecovered from combinatorial libraries and removed a significant bottleneck in the generation of affinityreagents on a proteomic scale.