Nebulabio is a protein expression team specializing in recombinant protein production using yeast-based eukaryotic platforms. By integrating microbial scalability with eukaryotic processing capacity, we support projects requiring reliable access to folded and, when applicable, secreted proteins within a cost-effective production framework. Our service focuses on delivering well-characterized protein material with traceable documentation for research and early development contexts.
☞Nebulabio’s Membrane Protein Platform
Membrane proteins are embedded in lipid bilayers and present unique physicochemical constraints, including extensive hydrophobic surfaces and instability upon delipidation. A successful production strategy therefore requires coordinated design across construct architecture, expression host, membrane extraction chemistry, and stabilization in a suitable membrane mimetic environment. Our platform integrates parallel construct evaluation, host selection across microbial and eukaryotic systems, detergent and lipid additive matrices, and optional reconstitution into nanodiscs or other mimetics, enabling delivery of monodisperse and structurally relevant membrane protein preparations.
| ☎ +86-15801534258 | ✉ info@nebulabio.cn | Contact Us ☜☜ |
☞Supported Options
Expression Hosts
• Prokaryotic hosts including E. coli with toxicity-mitigating strains such as C41 C43 and Lemo21
• Baculovirus insect cell systems including Sf9 Sf21 and High Five
• Yeast hosts including Pichia pastoris and Saccharomyces cerevisiae
• Mammalian hosts including HEK293 and CHO supporting transient and stable expression contexts
Construct Engineering
• Truncation and fusion partner panels including BRIL and T4 lysozyme designs when appropriate
• Targeted stabilizing mutation strategies for improved conformational stability
• Affinity tag placement and optimization including His FLAG and Strep tag II with function-aware configurations
Solubilization and Stabilization
• Detergent panels including DDM LMNG CHS OG and defined mixtures
• Lipid and additive supplementation including cholesterol and native lipid extracts when configured
• Membrane mimetics including nanodiscs amphipols and SMA-based systems as project-defined options
Purification and Characterization
• Multi-step chromatography including IMAC affinity capture and SEC polishing
• Homogeneity and oligomeric state assessment including SEC MALS when configured
• Thermal stability evaluation including DSF TSA and time-dependent stability tracking
• Functional evidence generation including ligand binding kinetics SPR BLI and transport related readouts when configured
☞Principle of Membrane Protein Production
Integral membrane proteins require a production strategy that respects their dependence on a lipid bilayer for structural stability. Construct selection typically begins with sequence-informed design to reduce flexible regions while preserving functional domains, followed by expression in a host that supports folding and membrane insertion. After membrane isolation, mild detergents are applied to extract the target while minimizing denaturation, and stabilization is further improved by lipid additives or reconstitution into membrane mimetics such as nanodiscs. Purification is performed under conditions that preserve monodispersity and functional state, enabling downstream use in structural determination, ligand interaction analysis, and mechanistic studies.
☞Nebulabio Service Workflow
Bioinformatics and Construct Design We establish a construct panel designed for stability expression feasibility and functional interpretability across selected hosts. Key Steps: • Target analysis including transmembrane prediction topology mapping and flexible region identification • Construct panel design including truncation variants fusion partner configurations and tag placements • Gene synthesis readiness review and cloning plan aligned with candidate host systems |
Small Scale Expression and Solubilization Screening We identify productive construct host and extraction chemistry combinations before committing to large-scale production. Key Steps: • Parallel expression across selected hosts and construct variants • Membrane fraction preparation and enrichment • Detergent matrix evaluation for extraction efficiency and stability behavior • Analytical confirmation including SDS PAGE and Western blot where configured |
Scale Up Expression and Membrane Preparation We scale the selected route and generate membrane material under controlled processing conditions. Key Steps: • Culture scale-up in flasks or bioreactors depending on host system • Cell harvest and controlled disruption • Differential centrifugation and ultracentrifugation for membrane isolation |
Solubilization Purification and Stabilization We extract the target protein under defined mild conditions and purify to a monodisperse preparation with stabilization options. Key Steps: • Solubilization using the selected detergent and additive context • Affinity capture and intermediate purification steps • Optional tag removal and cleanup • SEC polishing to obtain a homogeneous sample • Optional reconstitution into nanodiscs amphipols or SMA systems |
Functional and Biophysical Characterization We generate evidence supporting purity homogeneity stability and project-aligned functional state. Key Steps: • Purity and monodispersity evaluation including SEC UV and SEC MALS when configured • Stability assessment using DSF TSA and time-dependent monitoring • Functional evidence generation using ligand binding and project-defined activity assays |
Delivery and Documentation We deliver protein material and documentation enabling reproducible downstream handling and interpretation. Deliverables: • Purified stabilized membrane protein aliquots in detergent or reconstituted format as configured • COA including buffer detergent concentration purity and stability evidence • Full data package and project report with handling guidance |
☞Featured Applications of Nebulabio’s Services
● Structural Biology and Structure Enabled Discovery
We provide monodisperse membrane protein preparations suitable for crystallography and single particle cryo EM workflows, where sample stability and conformational homogeneity determine data quality. Reconstituted formats such as nanodiscs are used to preserve native-like conformations and support interpretable particle behavior in structural pipelines.
● High Throughput Screening and Lead Optimization
Purified functional targets enable biochemical or cell-free binding formats for compound screening and comparative ligand profiling. Defined protein preparations reduce variability introduced by heterogeneous membrane fractions and improve interpretability in kinetic and competition assays.
● Antibody Discovery against Membrane Targets
Native-like antigens in detergent micelles or nanodiscs support selection strategies that depend on correct folding and epitope presentation. This enables generation of binders recognizing conformational epitopes and improves downstream translation into functional blocking formats.
● Biosensor and Surface Immobilization Formats
Stabilized membrane proteins can be immobilized for label-free interaction measurements and biosensor formats when conformational integrity is preserved. Defined formulation and stability evidence support reproducible chip loading and comparative kinetic interpretation.
For more information, please contact us at info@nebulabio.cn or +86-15801534258.
