High-throughput screening of single-chain antibodies using multiplexed flow cytometry.

J Proteome Res

Bioscience Division, Mail Stop M888, Los Alamos National Laboratory, P.O. Box 1663, Los Alamos, New Mexico 87545, USA.

Published: March 2007

AI Article Synopsis

  • The new screening method utilizes multiplexed flow cytometry to efficiently analyze multiple affinity reagents at once for proteomic research, assessing expression levels and specific binding characteristics.
  • This technique provides higher quality and quantity of data with much faster analysis times and less antigen usage compared to traditional ELISA methods, collecting important information that is typically gathered in later screening stages.
  • By integrating high-throughput and multiplex technologies, the method streamlines the initial identification of affinity reagents from combinatorial libraries, addressing a major bottleneck in the process of generating these reagents on a large scale.

Article Abstract

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

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http://dx.doi.org/10.1021/pr0604108DOI Listing

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