Efficient clinical-grade γ-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing

γ-Retroviral vectors (γ-RV) are powerful tools for gene therapy applications. Current clinical vectors are produced from stable producer cell lines which require minimal further downstream processing, while purification schemes for γ-RV produced by transient transfection have not been thoroughly investigated. We aimed to develop a method to purify transiently produced γ-RV for early clinical studies. Here, we report a simple one-step purification method by high-speed centrifugation for γ-RV produced by transient transfection for clinical application. High-speed centrifugation enabled the concentration of viral titers in the range of 107-108 TU/mL with >80% overall recovery. Analysis of research-grade concentrated vector revealed sufficient reduction in product- and process-related impurities. Furthermore, product characterization of clinical-grade γ-RV by BioReliance demonstrated two-logs lower impurities per transducing unit compared with regulatory authority-approved stable producer cell line vector for clinical application. In terms of CAR T cell manufacturing, clinical-grade γ-RV produced by transient transfection and purified by high-speed centrifugation was similar to γ-RV produced from a clinical-grade stable producer cell line. This method will be of value for studies using γ-RV to bridge vector supply between early- and late-stage clinical trials

International relations in the making of political Islam: interrogating Khomeini's ‘Islamic government’

Eurocentric approaches to political Islam tend to deploy an internalist methodology that theoretically obscures the generative and constitutive role of international relations. This article addresses this problem through a critical application of Leon Trotsky's idea of ‘uneven and combined development’ to Ayatollah Khomeini's invention of the concept of ‘Islamic government’. It argues that this concept was international in its socio-political stimulus and intellectual content, and, crucially, reflected, influenced, and mobilised an emergent liminal sociality that combined Western and Islamic socio-cultural forms. This heterogeneous character of Iran's experience of modernity is, the article argues, theoretically inaccessible to Eurocentric approaches’ homogeneous and unilinear conceptions of history, which, as a result, generate exceptionalist modes of explanations