Multispecific Antibody Assembly via Sortase A Enzymatic Conjugation

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Solution Overview

Problem

Current therapeutic antibodies often have limited specificity and efficacy due to their inability to tailor their targeting and delivery to individual patient characteristics, leading to potential side effects and reduced effectiveness in treating diseases like cancer.

Innovation Solution

A method using Sortase A-mediated enzymatic conjugation to create tailor-made, multispecific antibodies that can be specifically designed for individual patients by determining their unique cell surface markers and combining antibody fragments with Sortase A, enabling improved targeting and delivery to specific antigens or epitopes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If conventional monoclonal antibodies are used for therapy, then they can provide immune-mediated effector functions, but they have limited specificity and cannot be tailored to individual patient characteristics

Engineering Contradiction:

ImprovespecificityVSAvoidcomplexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The antibody is divided into separate functional modules: Fv domains for antigen binding specificity and Fc regions for effector functions. This segmentation allows independent selection and combination of different antibody fragments to create multispecific entities tailored to specific patient needs while maintaining manageable complexity through modular assembly

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The Fc region serves multiple functions including mediating effector functions (ADCC, CDC, ADCP), interacting with FcRn for half-life extension, and providing a universal platform that can be combined with various Fv domains. This multi-functionality reduces the need for separate components and simplifies the overall design of personalized therapeutic antibodies

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Reliability

If monoclonal antibodies are designed to bind single antigens, then they have simple structure, but they have reduced effectiveness in treating complex diseases

Engineering Contradiction:

ImproveefficacyVSAvoidcomplexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

Multiple Fv domains with different antigen specificities are merged into a single multispecific antibody entity that can simultaneously bind multiple antigens or epitopes. This merging increases therapeutic efficacy by enabling coordinated targeting of multiple disease-relevant antigens while the Fc region provides unified effector function mediation

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The antibody is constructed as a composite molecule combining different antibody fragments (Fv domains from different sources or specificities) with an Fc region. This composite structure integrates diverse binding specificities with standardized effector functions, enhancing effectiveness against complex diseases while maintaining structural organization

Inventive Principle:

Principle #40Composite materials

3Reliability

If therapeutic antibodies are administered, then they can treat diseases, but they may cause side effects due to off-target binding

Engineering Contradiction:

ImproveselectivityVSAvoidside effects

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

Each Fv domain in the multispecific antibody is engineered with specific antigen binding specificity tailored to target only desired antigens present on disease-related cells. This localized optimization of binding properties at each antigen-binding site ensures high selectivity for target cells while minimizing off-target binding and associated side effects

Inventive Principle:

Principle #3Local quality

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

The approach results in highly specific, effective therapeutic molecules with reduced side effects by enhancing targeting and delivery, achieving improved selectivity and specificity through simultaneous binding to multiple epitopes, thereby increasing the therapeutic efficacy while minimizing off-target binding.

Implementation Method 1

A method using Sortase A-mediated enzymatic conjugation to create tailor-made, multispecific antibodies

Methodology Applied

Scientific EffectEnzymatic conjugation: Enzyme

Data Source

PatentUS20230220110A1Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof

Publication Date: 2023.07.13 F HOFFMANN LA ROCHE INC

AI summary

Herein is reported a method for producing a bispecific antibody comprising the step of incubating(i) an antibody Fab fragment or a scFv antibody comprising within the 20 C-terminal amino acid residues the amino acid sequence LPX1TG (SEQ ID NO: 01),(ii) a one-armed antibody comprising a full length antibody heavy chain, a full length antibody light chain, and an Fc-heavy chain,whereby the full length antibody heavy chain and the full length antibody light chain are cognate antibody chains that thereof forms an antigen binding site,whereby the full length antibody heavy chain and the Fc-heavy chain are covalently linked to each other via one or more disulfide bonds forming an antibody hinge region, andwhereby the Fc-heavy chain has an oligoglycine amino acid sequence at its N-terminus,and(iii) a Sortase A enzyme.