Recombinant CD40 Protein for B Cell Clonal Expansion
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Solution Overview
Problem
Current methods for generating fully human monoclonal antibodies are time-consuming and costly, and rodent antibodies are immunogenic in humans, limiting their use in immunotherapy.
Innovation Solution
A recombinant protein comprising an intracellular segment of CD40 and an amino acid sequence mediating association with the constant region of an immunoglobulin heavy chain, linked via a transmembrane domain, is used to induce clonal expansion and antibody secretion in B cells without T cell co-stimulation, allowing for the generation of antigen-specific antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional hybridoma or phage display techniques are used to generate monoclonal antibodies, then fully human antibodies can be produced, but the process is time-consuming and cost-intensive
Solution Approach 1:
The patent applies preliminary action by pre-transfecting B cells with the recombinant protein before antigen stimulation. This pre-preparation of B cells with the CD40-recombinant protein fusion enables them to respond more rapidly and efficiently to antigen challenge, skipping time-consuming steps in the traditional antibody generation process while maintaining the ability to produce fully human monoclonal antibodies
Solution Approach 2:
The patent uses copying by creating a recombinant protein that copies the essential signaling function of CD40 without requiring T cell co-stimulation. The recombinant protein fusion molecule mimics the CD40 receptor's ability to activate B cells, providing a simplified system that replicates the necessary biological activity in a more efficient manner
2Ease of manufacture
If rodent monoclonal antibodies are used for immunotherapy, then antibody-based therapy can be provided, but strong immunogenicity occurs in humans
Solution Approach 1:
The patent applies parameter changes by altering the species origin parameter of the antibody from rodent to human. By using human B cells as the source for antibody production and stimulating them with human recombinant protein, the resulting monoclonal antibodies have human sequence parameters that eliminate or reduce immunogenicity in human patients while maintaining therapeutic effectiveness
3Reliability
If T cell co-stimulation is used for B cell activation, then complete B cell activation and antibody secretion occur, but the process becomes more complex and requires additional cells
Solution Approach 1:
The patent extracts the essential CD40 signaling function from the complex T cell-B cell interaction system. By isolating and using the recombinant CD40 protein fused to the immunoglobulin constant region, the system removes the requirement for T cells and CD40L co-stimulation, simplifying the activation process while maintaining reliable B cell activation and antibody secretion
Solution Approach 2:
The recombinant protein fusion acts as an intermediary that bridges the gap between antigen recognition and B cell activation. The fusion molecule combines the immunoglobulin constant region (providing antigen binding) with the CD40 intracellular segment (providing activation signaling), eliminating the need for separate T cell intermediary components
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
This approach enables the efficient and cost-effective production of fully human monoclonal antibodies with predetermined antigen-specificity, suitable for immunotherapy, by mimicking the CD40 signal for B cell activation and differentiation.
Implementation Method 1
the amino acid sequence of an intracellular segment of CD40 or of a variant thereof which is capable of mediating the intracellular CD40 signal transduction
Implementation Method 2
an amino acid sequence mediating the association of the recombinant protein with the constant region of an immunoglobulin heavy chain
Implementation Method 3
an amino acid sequence comprising the amino acid sequence of a transmembrane domain
Implementation Method 4
inducing clonal expansion of B cells having a desired antigen-specificity
Data Source
AI summary
The present invention provides recombinant proteins comprising the amino acid sequence of an intracellular segment of CD40 and an amino acid sequence mediating the association of the recombinant protein with the constant region of an immunoglobulin heavy chain. The recombinant proteins according to the present invention are - useful for inducing clonal expansion of a B cell having a predetermined antigen-specificity without the need for T cell or CD40L mediated co-stimulation. Thus, the present invention provides tools for clonal expansion of B cells specific for an antigen of interest and the production of B cells secreting antibodies specific for an antigen of interest. The recombinant proteins of the present invention may also be used for generating fully human monoclonal antibodies with a predetermined antigen-specificity from the B cell repertoire of a human subject.