Antigen Binding Domain Linker Clipping Reduction
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Solution Overview
Problem
Biomolecules, particularly polypeptides and polypeptide constructs with antibody-derived binding domains, face significant stability issues due to clipping during liquid storage, which complicates pharmaceutical formulations and increases production costs, especially in regulated environments where stability and uniformity are crucial.
Innovation Solution
A polypeptide construct with a VH and VL variable region linked by a peptide linker comprising S(G4X)n or (G4X)n, where X is selected from Q, T, N, C, G, A, V, I, L, and M, and n is an integer from 1 to 20, is used to reduce clipping rates, replacing traditional S(G4S)n linkers and enhancing stability through conservative substitutions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If traditional S(G4S)n linkers are used to connect VH and VL variable regions, then the polypeptide construct can be manufactured with standard procedures, but the clipping rate increases during liquid storage reducing stability
Solution Approach 1:
The patent applies parameter changes by substituting specific amino acid residues in the peptide linker sequence. The traditional S(G4S)n linker is modified by replacing Serine (S) residues at positions 2 and 5 with alternative amino acids (X) selected from Q, T, N, C, G, A, V, I, L, or M, creating S(G4X)n or (G4X)n linkers. This sequence modification changes the chemical properties of the linker, reducing its susceptibility to clipping during liquid storage while maintaining the functional integrity of the polypeptide construct.
2Stability of the object's composition
If lyophilization is used to reduce clipping rate, then stability of biomolecule is improved, but production flexibility is reduced and cost of goods increases
Solution Approach 1:
The patent modifies the amino acid sequence parameters of the peptide linker to inherently reduce clipping susceptibility. By changing the linker composition to S(G4X)n or (G4X)n with specific amino acid substitutions, the biomolecule achieves improved stability during liquid storage without requiring lyophilization. This allows the biomolecule to be manufactured and stored in liquid form, maintaining production flexibility and reducing manufacturing complexity while achieving the desired stability improvement.
3Ease of operation
If liquid formulation is used for biomolecules, then handling safety and comfort are improved, but clipping occurs during storage reducing stability
Solution Approach 1:
The patent changes the amino acid composition parameters of the peptide linker by substituting Serine residues with alternative amino acids (X) from the specified group. This sequence modification reduces the linker's susceptibility to clipping during liquid storage, enabling the biomolecule to be formulated in liquid state. The modified S(G4X)n or (G4X)n linker maintains structural integrity during storage while allowing the biomolecule to retain the advantages of liquid formulation, including improved handling safety and comfort.
Data Source
AI summary
The invention relates to a polypeptide or polypeptide construct comprising a first target antigen binding domain, wherein said first target antigen binding domain comprises a VH and a VL variable region linked by a peptide linker, wherein the peptide linker comprises or consists of S(G4X)n or (G4X)n, wherein X is selected from the group consisting of Q, T, N, C, G, A, V, I, L, and M, and wherein n is an integer selected from integers 1 to 20. The invention also relates to a method for improving stability of a polypeptide or polypeptide construct. Moreover, the invention relates to a polynucleotide encoding the polypeptide or polypeptide construct of the invention, a vector comprising said polynucleotide and a host cell transformed or transfected with said polynucleotide or with said vector. Moreover, the invention also provides for a process for the production of said polypeptide or polypeptide construct and a pharmaceutical composition comprising said polypeptide or polypeptide construct of the invention. Furthermore, the invention relates to medical uses of said polypeptide or polypeptide construct and kits comprising said polypeptide or polypeptide construct.