Mutated Albumin Variants for Soluble Bacterial Expression
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Solution Overview
Problem
Existing bacterial expression systems face challenges in efficiently producing recombinant albumin proteins due to their large size and complex multidomain organization, leading to issues such as low solubility and stability, which are exacerbated by batch-to-batch variability and ethical concerns with animal-derived albumin.
Innovation Solution
Development of albumin protein variants with specific mutations, such as HSA1 and BSA1, that enhance thermostability and solubility, maintaining ligand binding affinity and increasing yield when expressed in bacteria, using a nucleic acid sequence encoding these variants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If albumin is expressed in bacterial systems, then production yield and scalability are improved, but solubility and stability deteriorate due to large size and complex multidomain organization
Solution Approach 1:
The patent applies parameter changes by systematically mutating amino acid residues at protein-protein interface regions and hydrophobic core regions of albumin. Specific substitutions (e.g., L198H, V120P, F156Y, D187E, L198H, S202I, V310I, A371S, V381I, V409I, S427A, V455I, K519E, A552S, V576I) were introduced to alter the physical-chemical parameters of the protein, thereby improving solubility and stability while maintaining functionality in bacterial expression systems.
2Reliability
If animal-derived albumin is used, then functional performance is maintained, but ethical concerns and batch-to-batch variability increase
Solution Approach 1:
The patent creates recombinant albumin variants that copy the essential functional properties of native animal-derived albumin through genetic engineering. By synthesizing the albumin gene sequence and expressing it in bacterial systems with optimized mutations, the invention produces a bacterial-derived copy that maintains ligand binding affinity and structural integrity while eliminating ethical concerns and batch variability associated with animal sourcing.
3Ease of manufacture
If wild type albumin is expressed in bacteria, then production is attempted, but solubility and stability are insufficient for efficient production
Solution Approach 1:
The patent applies preliminary action by pre-optimizing the albumin sequence through in silico mutagenesis and computational modeling before actual bacterial expression. The amino acid mutations were designed and validated in silico to predict improved solubility and stability, allowing the production process to start with an already optimized sequence rather than attempting to express wild type albumin and then troubleshoot solubility issues.
Data Source
AI summary
An albumin protein is provided. The albumin protein is soluble when expressed in bacteria and characterized by increased thermostability as compared to wild type human serum albumin (HSA). Also provided are nucleic acid molecules encoding the albumin protein, compositions comprising the albumin protein and uses thereof.