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 Expression and Purification Platform
Our integrated platform supports parallel construct panels, host evaluation across microbial and eukaryotic systems, high-coverage detergent and additive matrices, and optional reconstitution into nanodiscs or detergent-free mimetics. This enables delivery of purified preparations with defined formulation and analytical evidence supporting stability and homogeneity.
| ☎ +86-15801534258 | ✉ info@nebulabio.cn | Contact Us ☜☜ |
☞Supported Options
Expression Host Systems
• E. coli with specialized strains including C41 C43 and Lemo21 for toxicity mitigation
• Baculovirus insect cell systems including Sf9 and High Five for complex multi-pass targets
• Yeast systems including Pichia pastoris and Saccharomyces cerevisiae for scalable eukaryotic expression
• Mammalian systems including HEK293 and CHO supporting transient and stable expression contexts
Construct Design and Engineering
• Truncation and fusion partner panels including BRIL T4 lysozyme and MBP where appropriate
• Stabilizing mutation strategies to enhance conformational stability
• Affinity tag optimization including His FLAG and Strep tag II with function-aware placement
Solubilization and Stabilization
• High-coverage detergent matrices including DDM LMNG CHAPS OG and defined mixtures
• Lipid and additive supplementation including cholesterol and native lipid extracts when configured
• Reconstitution options including nanodiscs amphipols and SMA-based formats for detergent-free stabilization
Purification and Quality Control
• Multi-step chromatography including affinity capture and SEC polishing
• Homogeneity and aggregation assessment including SEC MALS when configured
• Stability assessment including DSF TSA and time-dependent monitoring
• Functional validation options including SPR BLI fluorescence-based activity and transport assays as configured
☞Principle of Membrane Protein Expression and Purification
Membrane proteins require a production strategy that preserves their native-like conformational state when removed from the lipid bilayer. Construct design typically aims to reduce conformational heterogeneity while maintaining functional domains, often via rational truncations, fusion partners, and stabilizing mutations. Expression is performed in a host that supports membrane insertion and folding, after which membranes are isolated and the target is extracted using mild detergents that substitute for native lipids while minimizing denaturation. Purification proceeds in detergent-containing buffers or through reconstitution into membrane mimetics such as nanodiscs, yielding monodisperse preparations suitable for structural analysis, ligand interaction studies, and mechanistic assays.
☞Nebulabio Service Workflow
Bioinformatics and Construct Design We establish a construct panel designed for expression feasibility stability and downstream interpretability. Key Steps • Topology and transmembrane prediction with domain boundary mapping • Construct panel design including truncations fusion partners and affinity tag configurations • Gene readiness review and cloning plan aligned with candidate host systems |
Small Scale Expression and Solubilization Screening We identify productive host construct and extraction condition combinations before scale-up. Key Steps • Parallel expression across selected hosts and construct variants • Membrane enrichment and pilot extraction using detergent matrices • Analytical confirmation including SDS PAGE and Western blot where configured • Preliminary stability behavior evaluation to guide scale-up selection |
Large Scale 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 • Controlled harvest and disruption with membrane fraction isolation • Crude membrane preparation suitable for reproducible extraction |
Solubilization Purification and Reconstitution We extract the target under defined mild conditions and purify to a homogeneous preparation with optional reconstitution. Key Steps • Optimized detergent extraction with selected lipid additive context • Affinity capture and intermediate purification steps • Optional tag cleavage and cleanup • SEC polishing to obtain monodisperse sample • Optional reconstitution into nanodiscs amphipols or SMA formats |
Functional and Biophysical Characterization We generate evidence supporting purity homogeneity stability and functional state aligned to project endpoints. Key Steps • Purity and homogeneity assessment including SEC UV and SEC MALS when configured • Stability assessment using DSF TSA and time-dependent monitoring • Functional validation using ligand binding kinetics SPR BLI and project-defined activity assays |
Delivery and Documentation We deliver protein material and documentation enabling reproducible downstream handling and interpretation. Deliverables • Aliquoted membrane protein in defined detergent or reconstituted format with specified storage condition • COA including formulation concentration purity and stability evidence • Full data package and project report including handling and thawing guidance |
☞Featured Applications of Nebulabio’s Services
● Structural Biology and Cryo EM
Purified monodisperse preparations are supplied for crystallography and single particle cryo EM where sample stability and conformational homogeneity determine achievable resolution. Reconstituted formats such as nanodiscs can preserve native-like constraints and improve interpretability of particle populations during structure determination.
● High Throughput Drug Screening
Functional purified GPCRs channels and transporters support biochemical and cell-free binding formats for compound library screening and comparative ligand profiling. Defined preparations reduce variability associated with heterogeneous membrane fractions and strengthen kinetic interpretation in competition and affinity assays.
● Therapeutic Antibody Discovery
Native-like membrane protein preparations in detergents or nanodiscs provide antigens with conformational epitope preservation, supporting immunization or selection workflows that depend on correct folding. This improves the likelihood of obtaining binders recognizing functional states rather than denatured surfaces.
● Mechanistic and Transport Studies
Reconstitution into proteoliposomes or nanodiscs enables controlled assays of transport flux gating or inhibition under defined lipid contexts. This supports comparative kinetic analysis across variants and provides interpretable inhibition profiles for mechanistic evaluation.
For more information, please contact us at info@nebulabio.cn or +86-15801534258.
