Multiparameter screen optimizes immunoprecipitation

Biotechniques. 2024 Feb 29. doi: 10.2144/btn-2023-0051. Online ahead of print.ABSTRACTImmunoprecipitation (IP) coupled with mass spectrometry effectively maps protein-protein interactions when genome-wide, affinity-tagged cell collections are used. Such studies have recorded significant portions of the compositions of physiological protein complexes, providing draft 'interactomes'; yet many constituents of protein complexes still remain uncharted. This gap exists partly because high-throughput approaches cannot optimize each IP. A key challenge for IP optimization is stabilizing in vivo interactions during the transfer from cells to test tubes; failure to do so leads to the loss of genuine interactions during the IP and subsequent failure to detect. Our high-content screening method explores the relationship between in vitro chemical conditions and IP outcomes, enabling rapid empirical optimization of conditions for capturing target macromolecular assemblies.PMID:38425263 | DOI:10.2144/btn-2023-0051

Source: BioTechniques - March 1, 2024 Category: Biotechnology Authors: Shaoshuai Xie Leila Saba Hua Jiang Omar R Bringas Mehrnoosh Oghbaie Luciano Di Stefano Vadim Sherman John LaCava Source Type: research