Humanized ISVDs for CTLA-4 Binding with Reduced Immunogenicity
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Solution Overview
Problem
Current methods for inhibiting CTLA-4 mediated downregulation, such as using Nanobodies, face challenges due to potential unwanted anti-drug immune responses, necessitating the development of novel humanized Nanobodies that minimize pre-existing antibody binding.
Innovation Solution
Development of multispecific immunoglobulin single variable domains (ISVDs) that bind to human CTLA-4 at specific residues, including VRVTVL, ADSQVTEVC, and CKVELMYPPPYYLG, with mutations at positions 11, 89, and 112 to reduce pre-existing antibody binding, and are designed to prevent CTLA-4 from binding to CD80 and CD86, thereby blocking immune tolerance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Nanobodies are used to inhibit CTLA-4, then immune suppression is blocked and T-cell activation is enhanced, but pre-existing antibodies in patient serum bind to the Nanobodies causing unwanted anti-drug immune responses
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the Nanobody at specific positions (11, 89, and 112 according to Kabat numbering). These sequence modifications alter the molecular structure to reduce recognition and binding by pre-existing human antibodies, thereby decreasing immunogenicity while preserving the ability to bind CTLA-4 and block immune suppression.
2Object-affected harmful factors
If the Nanobody structure is modified to reduce pre-existing antibody binding, then anti-drug immune response is minimized, but binding affinity to CTLA-4 may be affected
Solution Approach 1:
The patent applies local quality by making targeted modifications at specific local positions (residues 11, 89, and 112) of the Nanobody sequence rather than global changes. These localized sequence changes are strategically selected to reduce pre-existing antibody binding while preserving the critical binding interface with CTLA-4, thus maintaining binding affinity while reducing immunogenicity.
Data Source
AI summary
The present invention provides molecules, such as ISVDs and Nanobodies, that bind to CTLA4 or human serum albumin. These molecules have been engineered so as to reduce the incidence of binding by pre-existing antibodies in the bodies of a subject administered such a molecule. Methods for increasing immune response, treating cancer and/or treating an infectious disease with such molecules are provided.