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 Yeast Expression Platform
Yeast expression combines the operational simplicity of microbial fermentation with eukaryotic protein processing, supporting disulfide bond formation, proteolytic processing, and N- and O-linked glycosylation patterns that differ from mammalian profiles. Our platform covers multiple yeast hosts and provides an end-to-end workflow from gene and cassette strategy through strain construction, expression screening, fermentation execution, purification, and characterization. The service supports intracellular and secreted formats across research-scale production and pilot-scale runs with defined milestones to maintain consistency across batches.
| ☎ +86-15801534258 | ✉ info@nebulabio.cn | Contact Us ☜☜ |
☞Supported Options
Yeast Hosts
• Saccharomyces cerevisiae
• Pichia pastoris and Komagataella phaffii
• Kluyveromyces lactis
• Custom-engineered strains including glycoengineered or protease-deficient variants
Vectors and Promoters
• Inducible systems including AOX1 GAL1 10 and CUP1
• Constitutive systems including GAP TEF1 and PGK1
• Secretion leaders including alpha factor and host-adapted secretion signals
• Tags and fusion partners including His FLAG c Myc GST Fc and solubility partners
Fermentation and Process Modes
• Clone and condition screening in shake flasks
• High-cell-density fed-batch in bioreactors
• Methanol-inducible or methanol-free routes depending on host system
• Extended production modes including continuous or perfusion-like operation where applicable
Downstream Processing
• Cell disruption routes for intracellular targets
• Clarification and filtration for secreted products
• Affinity capture and chromatography polishing including ion exchange hydrophobic interaction and size exclusion
• Buffer exchange and concentration into defined formulations
☞Principle of Yeast Expression Systems
Yeast are unicellular eukaryotes that combine rapid microbial growth with core eukaryotic processing functions. Recombinant genes are maintained episomally or integrated into the genome and expressed under strong yeast promoters in either inducible or constitutive modes. Yeast can support disulfide bond formation, proteolytic processing, and N- and O-linked glycosylation, although glycan structures can differ substantially from mammalian patterns. This makes yeast platforms broadly suitable for enzymes, secreted proteins, antigens, and complex targets that benefit from eukaryotic folding while remaining compatible with yeast-specific processing outcomes.
☞Nebulabio Service Workflow
Project Design and Strain Engineering We define the host and cassette strategy to match target class secretion intent and scale expectations, then generate expression strains for screening. Key Steps • Host selection aligned with target attributes and secretion intent • Cassette design with promoter leader tag and selection marker logic • Gene readiness review and codon adaptation for the chosen yeast context • Transformation and preliminary clone identification for downstream screening |
Small Scale Expression and Screening We evaluate expression behavior across candidate clones and establish a condition set suitable for production. Key Steps • Clone evaluation in deep-well plates or shake flasks • Induction or constitutive expression evaluation across defined time windows • Secreted product assessment from clarified supernatant or intracellular assessment from lysate • Analytical confirmation using SDS PAGE and Western blot when configured |
Process Development and Fermentation Scale Up We transition from screening to controlled production using a defined inoculum train and monitored fermentation execution. Key Steps • Inoculum train development from starter culture to seed stage • Fed-batch fermentation execution with controlled carbon source strategy • Monitoring of DO pH and temperature during the production phase • Harvest definition aligned with secreted or intracellular product format |
Purification and Polishing We recover target protein and apply chromatography steps to reach the agreed purity and homogeneity tier. Key Steps • Supernatant clarification and filtration for secreted formats • Cell disruption and debris removal for intracellular formats • Affinity capture followed by ion exchange hydrophobic interaction or size exclusion polishing as configured • Concentration and buffer exchange into storage or assay conditions |
Quality Control and Characterization We generate a traceable characterization package supporting identity purity processing state and project-defined functional attributes. Key Assays • Purity and integrity assessment by SDS PAGE and SEC HPLC or CE SDS when configured • Concentration determination by UV280 BCA or ELISA where applicable • Identity support by peptide mapping or intact mass LC MS when configured • Glycosylation profiling by lectin blot or MS-based glycan analysis when configured • Activity assays aligned with project-defined endpoints |
Delivery and Documentation We deliver aliquoted protein material with complete documentation to support reproducible downstream use. Deliverables • Purified protein aliquots with defined storage condition • COA and QC summary with associated raw data • Process summary aligned with fermentation and purification route when configured • Handling and storage guidance including reconstitution notes where relevant |
☞Featured Applications of Nebulabio’s Services
● Therapeutic Format Proteins and Antibody Fragments
Yeast expression supports scalable production of antibody fragments and secreted proteins where microbial fermentation economics and eukaryotic folding are jointly required. Documented purity and processing state enable comparability in variant evaluation and downstream assay work.
● Vaccine Antigens and VLPs
Yeast platforms are frequently used to generate recombinant antigens and self-assembling particle-like materials in contexts where consistent supply and defined composition are required. Characterization packages support lot comparability during iterative antigen format evaluation.
● Membrane Proteins and Receptor Expression
Yeast membranes provide a eukaryotic lipid environment that can support certain membrane targets and receptor domains, enabling materials for structural and ligand interaction studies. Project-defined workflows align expression format with downstream readouts.
● Protein Engineering and Library Screening
Yeast surface display provides a eukaryotic presentation context for binding and stability selection workflows. It supports comparative evaluation of variants under consistent display and detection conditions.
For more information, please contact us at info@nebulabio.cn or +86-15801534258.
