About Us

Nebulabio is a premier contract research organization (CRO) dedicated to accelerating therapeutic, diagnostic, and research breakthroughs through comprehensive monoclonal antibody (mAb) discovery and development services. With a deep commitment to scientific excellence and innovation, we partner with biotech companies, academic institutions, and pharmaceutical leaders worldwide to transform novel targets into high-quality, functional antibody candidates. Our team of experienced scientists leverages state-of-the-art platforms and a flexible, collaborative approach to navigate the complexities of antibody generation. We understand that each project is unique, and we tailor our strategies to meet specific requirements for affinity, specificity, species, format, and intended application. From target validation to purified lead candidate, we provide an integrated, seamless pipeline designed to reduce timelines and mitigate risks, empowering our clients to advance their programs with confidence.

☞Nebulabio’s Monoclonal Antibody Discovery Workflow

Our systematic, multi-stage workflow ensures the identification and characterization of optimal antibody leads.

①Project Initiation & Antigen Design: We collaborate closely with clients to define project goals. Our services include antigen preparation (recombinant proteins, peptides, cell lysates, or whole cells) and strategy design for immunization or library screening.

②Immunization & Immune Library Construction: For hybridoma projects, we employ optimized immunization protocols in host species (mouse, rat, rabbit) to generate a robust immune response. For antibody library approaches, we construct immune libraries from immunized hosts or synthetic/semi-synthetic naïve libraries.

③Antibody Generation:

• Hybridoma Technology: Spleen cells from immunized animals are fused with myeloma cells. The resulting hybridomas are screened to isolate clones producing target-specific antibodies.

• Phage Display Technology: Vast libraries of antibody fragments (scFv, Fab) displayed on phage particles are panned against the antigen to enrich binders over multiple rounds.

• Single B Cell Sorting: Using fluorescently labeled antigen, antigen-specific B cells are isolated via flow cytometry from immunized hosts, followed by direct amplification of antibody genes.

④Primary & Secondary Screening: High-throughput screening (ELISA, FACS, bio-layer interferometry) is performed to identify clones with desired binding characteristics to the target antigen and assess cross-reactivity.

⑤Lead Characterization: Positive hits undergo deep characterization, including:

• Sequence Analysis: VH/VL gene sequencing, germline analysis, and humanization design.

• Affinity Measurement: Determination of binding kinetics (KD, Kon, Koff) using SPR (Biacore) or BLI.

• Specificity & Cross-Reactivity: Profiling against related protein families or cell lines.

• Functional Assays: In vitro evaluation of neutralization, blocking, agonist/antagonist activity, or internalization.

⑥Antibody Engineering & Optimization: Services include humanization, affinity maturation, isotype switching, and generation of various formats (e.g., IgG, Fab, scFv, bispecifics).

⑦Recombinant Expression & Purification: Lead antibody genes are cloned into mammalian expression vectors (e.g., CHO, HEK293) for small-scale transient or stable production. Antibodies are purified via Protein A/G affinity chromatography, followed by polishing steps.

☞Technologies for Monoclonal Antibody Discovery and Development

Our integrated toolkit features industry-standard and cutting-edge platforms:

①Diverse Animal Hosts: We offer mouse, rat, and rabbit hybridoma generation. Rabbit antibodies are particularly valuable for targeting difficult antigens due to their high affinity and epitope diversity.

②Advanced Display Technologies:

• Phage Display: Robust platform for screening large combinatorial libraries (≥10^9 members) for binders.

• Yeast Surface Display: Enables quantitative screening and fine affinity discrimination using fluorescence-activated cell sorting (FACS).

• Mammalian Cell Surface Display: Facilitates direct discovery of antibodies with optimal expression and stability in mammalian systems.

③Single B Cell Cloning Technologies: This platform allows for the direct isolation of naturally paired, antigen-specific heavy and light chain genes, preserving the native immune repertoire.

④Nanobody Discovery: We provide specialized services for generating single-domain antibodies (VHHs) from immunized camelids or via library screening, ideal for targeting cryptic epitopes and for diagnostic imaging.

⑤High-Throughput Screening & Analytics: Automated liquid handling, multiplexed ELISA, flow cytometry, and real-time interaction analysis (SPR, BLI) enable rapid identification and validation of leads.

⑥Antibody Engineering & Humanization: Using CDR-grafting and framework optimization techniques to reduce immunogenicity for therapeutic development while maintaining binding and function.

Figure1.Overview of five key approaches for therapeutic antibody development: (a) mouse hybridoma, (b) phage display, (c) transgenic mouse, (d) single B cell isolation, and (e) de novo design. 

☞Applications of Monoclonal Antibodies

The mAbs discovered and developed through our services power advancements across multiple sectors:

☞Nebulabio’s Service Highlights & Advantages

• Integrated End-to-End Solutions: A single provider for the entire journey from antigen design to purified, characterized antibody, ensuring continuity and streamlined communication.

• Platform Flexibility & Expertise: No one-size-fits-all approach. We recommend and apply the most suitable technology (hybridoma, phage display, single B cell) based on your target and project goals.

• Proven Track Record & Experience: Our team has successfully delivered antibodies against a wide array of targets, including membrane proteins (GPCRs, ion channels), post-translationally modified antigens, and toxic proteins.

• High-Quality, Functional Outputs: We focus not just on binding, but on delivering antibodies with the functional characteristics required for your downstream applications, supported by comprehensive data packages.

• Rapid Timelines & Transparent Processes: Optimized workflows and parallel processing steps accelerate lead identification. Clients receive regular updates and have access to a dedicated project manager.

• Customization & Collaboration: We adapt our strategies to your specific needs, whether for research use, diagnostic development, or therapeutic lead generation, functioning as a true extension of your R&D team.

• Strong Intellectual Property (IP) Support: We operate under clear confidentiality agreements and can tailor work plans to support your IP strategy and freedom-to-operate.

By partnering with Nebulabio for your monoclonal antibody discovery needs, you leverage a combination of robust technology, deep scientific knowledge, and a client-focused partnership model designed to de-risk and accelerate your path to success.