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.

Twist’s Precision Variant Library Technology stands out as a groundbreaking approach for focused screening in protein engineering. Powered by Twist’s massively parallel silicon-based DNA synthesis platform, this technology delivers highly uniform and accurate oligos, with an impressive 90% of oligos represented within <2.5x of the mean. Additionally, it boasts an industry-leading low error rate of 1:2,000 nt.

The combination of this cutting-edge synthesis platform with Twist’s well-established molecular biology expertise allows the creation of highly diverse gene mutant libraries. These libraries exhibit excellent variant representation and offer highly specific user-defined compositions, free from unwanted bias or motifs. With precise control over codon usage, amino acid distribution, and length variation, Twist’s library technology enables comprehensive exploration of the variant sequence space. The key benefits include high diversity precision, verified quality through rigorous quality control and NGS verification, and unmatched flexibility in designing sequences, domains, and combinations. This technology outperforms alternatives like TRIM and the degenerate approach, providing researchers with a superior tool for efficient and tailored protein engineering studies.