HIV-1 gp140 Immunogen Stabilization via HR1 Loop and Disulfide Engineering
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Solution Overview
Problem
Current HIV-1 vaccine development faces challenges in generating an antibody response capable of neutralizing a broad range of clinical isolates due to the genetic diversity of HIV-1, leading to the failure of existing candidate vaccines in clinical testing.
Innovation Solution
The development of modified HIV-1 envelope gp140 proteins, which consist of a gp120 polypeptide and a gp41 polypeptide with a redesigned N-terminus of the heptad 1 region (HR1) stabilized by a loop sequence, along with a flexible linker sequence at the cleavage site and an engineered disulfide bond, to enhance trimer stability and immunogenicity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional gp140 proteins are used, then the structure is simpler to produce, but the trimer stability and immunogenicity are insufficient
Solution Approach 1:
The gp140 protein is divided into separate gp120 and gp41 polypeptide chains that are non-covalently associated, allowing independent optimization of each component's stability and immunogenicity while maintaining the functional trimeric structure
Solution Approach 2:
The patent creates a composite protein structure by non-covalently associating gp120 and gp41 polypeptides to form a stable trimeric complex, combining the antigenic properties of gp120 with the structural stability of gp41 to achieve both reliability and controlled complexity
2Stability of the object's composition
If the N-terminus of HR1 region is redesigned with loop sequence, then the pre-fusion structure is stabilized, but the manufacturing process becomes more complex
Solution Approach 1:
The patent applies localized structural modification by redesigning only the N-terminus of the HR1 region with a specific loop sequence, while leaving the rest of the gp140 structure unchanged, thereby achieving pre-fusion stabilization with minimal impact on overall manufacturability
Solution Approach 2:
The invention modifies specific structural parameters of the HR1 region by introducing a loop sequence of defined length and composition, changing the local conformational parameters to stabilize the pre-fusion state without requiring complete restructuring of the protein
3Productivity
If engineered disulfide bonds are introduced, then the trimer yield and purity improve, but the protein engineering complexity increases
Solution Approach 1:
The patent introduces engineered disulfide bonds at predetermined positions within the gp41 polypeptide before protein expression and assembly, pre-establishing stable structural anchors that guide proper trimer formation and enhance yield during the manufacturing process
Solution Approach 2:
The invention creates a composite structural framework by integrating disulfide cross-linked regions within gp41 that reinforce the trimeric assembly, combining covalent and non-covalent interactions to achieve high yield and purity
4Reliability
If flexible linker sequence is added at cleavage site, then the immunogenicity is enhanced, but the protein structure complexity increases
Solution Approach 1:
The patent introduces a flexible linker sequence as an intermediary element at the cleavage site between gp120 and gp41, allowing proper spatial arrangement and orientation of the two polypeptides while enhancing immunogenicity through increased conformational flexibility and epitope accessibility
Data Source
AI summary
The present invention provides HIV-1 vaccine immunogens. Some of the immunogens contain a soluble gp140-derived protein that harbors a modified N-terminus of the HR1 region in gp41. Some of the immunogens contain an HIV-1 Env-derived trimer protein that is presented on a nanoparticle platform. The invention also provides methods of using the HIV-1 vaccine immunogens for eliciting an immune response or treating HIV infections.