We have developed a rapid process for cell-free production of full-length ErbB2, a receptor tyrosine kinase that contributes to the onset and progression of breast and other tumor types. The receptor is solubilized and stabilized via incorporation into nanolipoprotein particles (NLPs). NLPs are discoidal structures, approximately 20 nm in diameter, that arise from the self-assembly of apolipoproteins around a phospholipid bilayer that acts as a mimic of the cell membrane. The ErbB2-NLP complex was soluble and could be recognized by antibodies, such as Trastuzumab, that indicate the presence of correctly folded ErbB2 extracellular domains. Kinase function was confirmed using Lapatinib, which inhibited tyrosine autophosphorylation during cell-free synthesis. The cell-free synthesis and biochemical characterization data support the potential use of ErbB2-NLPs and related variants for novel biochemistry and drug screening studies.

Citation Format: Wei He, Matthew Saldana, Tiffany Scharadin, Candice Gellner, Yuanpei Li, Kit Lam, Kermit Carraway, Paul Henderson, Matthew Coleman. Cell-free generation and characterization of ErbB2-nanoparticle conjugates. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5660. doi:10.1158/1538-7445.AM2013-5660