Fibronectin Scaffold Proteins Stability via Sequence Modification
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Solution Overview
Problem
Protein pharmaceuticals face physical and chemical instability during production, purification, storage, and delivery, leading to reduced efficacy due to fragmentation and aggregation issues.
Innovation Solution
Development of novel fibronectin-based scaffold proteins with enhanced stability, featuring a fibronectin type III tenth (10Fn3) domain with specific amino acid sequences and modifications, such as C-terminal tails and N-terminal extensions, that reduce fragmentation and aggregation, and are designed to bind target molecules with high affinity while avoiding binding to unwanted proteins like EGFR and human serum albumin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antibodies are used, then they can bind to target molecules, but they lose structure and function under certain conditions due to instability
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the fibronectin type III domain, specifically optimizing parameters such as hydrophobic core packing, salt bridge formation, and disulfide bond placement to enhance structural stability while maintaining binding function under challenging conditions
2Productivity
If protein pharmaceuticals are produced and stored, then they can be delivered therapeutically, but they undergo fragmentation and aggregation reducing efficacy
Solution Approach 1:
The patent extracts and eliminates problematic sequences from the fibronectin domain that predispose the protein to fragmentation and aggregation, specifically removing unstable regions while retaining the essential binding functionality and structural integrity
Solution Approach 2:
The patent creates a composite structure by combining stabilized fibronectin type III domains with specific C-terminal extensions and N-terminal sequences that collectively provide enhanced resistance to fragmentation and aggregation during production, storage, and delivery
3Measurement precision
If fibronectin scaffold proteins are designed for high affinity binding, then they can effectively bind target molecules, but they may bind to unwanted proteins reducing specificity
Solution Approach 1:
The patent applies local quality by making specific localized modifications to the loop regions (AB, CD, EF) of the fibronectin type III domain that are responsible for target binding, while keeping the rest of the structure stable and non-reactive, thereby achieving high affinity for the intended target while minimizing off-target binding
Data Source
AI summary
The present application provides fibronectin based scaffold proteins associated with improved stability. The application also relates to stable formulations of fibronectin based scaffold proteins and the use thereof in diagnostic, research and therapeutic applications. The application further relates to cells comprising such proteins, polynucleotides encoding such proteins or fragments thereof, and to vectors comprising such polynucleotides.