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CAR Library

CAR Library

In recent years, chimeric antigen receptor (CAR) T-cell therapy has emerged as a promising form of immunotherapy, offering new avenues for cancer treatment. Advances in gene transfer technology and gene editing, coupled with innovative CAR designs, have propelled the development of novel cancer therapies. However, despite these strides, several challenges remain, necessitating further research and development efforts to accelerate progress in the field.

CARs typically comprise an extracellular binding domain, a hinge region, a transmembrane domain, and one or more intracellular domains. Fine-tuning each component of the CAR is crucial for enhancing T cell specificity, antigen recognition, and overall T cell function. Studies have demonstrated that even minor modifications to the CAR can significantly impact therapeutic outcomes. Therefore, it is imperative to have a comprehensive library for thorough testing, considering the complexity of each CAR construct.

Twist has pioneered a groundbreaking technology for constructing CAR libraries, allowing seamless shuffling of variants within each domain through a scarless assembly process. The key benefits of this technology include high diversity and quality, with each combinatorial assembly accommodating up to 10,000 gene fragment combinations. NGS-verified libraries ensure that over 90% of possible variants are present within 10 times of the mean. The flexibility and customization options offered by Twist’s technology allow users to design TCR and CAR libraries with user-defined combinatorial variants across specific elements. Additionally, the platform provides flexibility in throughput, enabling the insertion of sequences up to 1.5 kb in length and designing libraries at scale with diversity across multiple elements of the sequence. Twist’s technology represents a significant advancement in the development and optimization of CAR T-cell therapies.

The process begins with synthesizing genes representing different sequence variants for each domain of the Chimeric Antigen Receptor (CAR). These domains, including the extracellular binding domain, hinge region, transmembrane domain, and intracellular domains, can consist of multiple sequence variations. Twist then combines these synthesized genes through a scarless assembly process. This unique assembly method ensures that the combination of these genes results in a highly diverse library for scaffold optimization.

The scarless assembly process enables the seamless merging of the gene variants, creating a comprehensive library that spans a wide range of potential combinations. This technology allows for the exploration of unique domain combinations, unveiling novel functionalities within the CAR scaffold.

The workflow for Chimeric Antigen Receptor (CAR) discovery and optimization involves studying how each domain of the CAR scaffold independently and synergistically influences its functionality. CAR Libraries, generated through this technology, act as a valuable tool for fine-tuning each module. This process helps uncover codependencies among the domains and provides a deeper understanding of their impact on T-cell specificity, antigen recognition, and overall T-cell function.

Researchers can partner with Twist at any stage of their discovery workflow, leveraging both in vivo and in vitro workflows for binder discovery and optimization. Additionally, the use of synthetic libraries from Twist enhances scaffold optimization and validation, providing a versatile and efficient approach to advancing CAR T-cell therapy research and development.

different-sequence-variants-scaffold-optimization
T Cell Receptor (TCR) Discovery workflow using Combinatorial Assembly Libraries
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