Sortase CAR T Cells for Site-Specific Surface Modification
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Solution Overview
Problem
Current methods lack effective solutions for site-specific modification of eukaryotic cells to attach moieties and functional groups to their surfaces, limiting the ability to alter virulence, infection, and colonization abilities of pathogens, and for targeted therapeutic applications such as cancer treatment.
Innovation Solution
Development of sortase immune receptors and chimeric antigen receptors (CARs) comprising a sortase enzymatic region, transmembrane domain, and intracellular domain, which recognize specific antigen-binding domains and motifs to mediate site-specific attachment of proteins and therapeutic agents to cell surfaces.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If genetic modification methods are used to alter cell surface proteins, then the ability to change virulence and infection ability is improved, but the precision and control over specific attachment sites is lost
Solution Approach 1:
The patent introduces a sortase enzyme as an intermediary that specifically recognizes and binds to the LPXTG motif on cell surface proteins. This enzyme mediates the attachment of therapeutic agents to precisely defined sites, combining the versatility of genetic modification with the precision of targeted attachment. The sortase enzyme acts as a bridge between the antigen-binding domain and the cell surface protein, enabling controlled conjugation at specific locations.
Solution Approach 2:
The invention implements local quality by creating a specific recognition motif (LPXTG) at the desired attachment site on the cell surface protein. This localized feature allows the sortase enzyme to selectively attach therapeutic agents only at this specific site, rather than throughout the entire protein structure. The antigen-binding domain is engineered with this specific local characteristic that enables precise targeting.
2Reliability
If multiple moieties are attached to cell surface proteins, then therapeutic efficacy is improved, but the complexity of controlling attachment stoichiometry increases
Solution Approach 1:
The patent applies preliminary action by pre-engineering the antigen-binding domain with the LPXTG recognition motif incorporated into its structure. This pre-prepared configuration allows the sortase enzyme to readily identify and attach multiple therapeutic agents to the same standardized site, simplifying the control of attachment stoichiometry. The motif is built in advance, eliminating the need for complex real-time control mechanisms during the attachment process.
Solution Approach 2:
The sortase enzyme provides universality by recognizing and binding to the same LPXTG motif regardless of which therapeutic agent is being attached. This universal recognition system allows multiple different moieties to be attached using the same enzymatic mechanism, simplifying the overall process. The enzyme can attach various therapeutic agents (cytotoxic drugs, radioactive isotopes, etc.) to the antigen-binding domain through a consistent, standardized pathway.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables targeted and specific attachment of therapeutic agents to cell surfaces, enhancing therapeutic efficacy in treating diseases like cancer and autoimmune disorders by allowing precise control over the attachment of moieties and functional groups.
Implementation Method 1
The sortase enzyme attaches to a 'sortase substrate motif ', such as Leu-Pro-any-Thr-Gly (LPXTG) (SEQ ID NO: 2), cleaves the peptide between the Thr and Gly, and then covalently attaches this protein to an acceptor peptide, such as polyglycine via a transpeptidation reaction.
Data Source
AI summary
The present invention includes compositions and methods comprising sortase immune receptors and sortase chimeric antigen receptors (CARs).