About Us

Nebulabio is a dedicated monoclonal antibody development team specializing in classic hybridoma technology, providing stable and scalable solutions for monoclonal antibody generation. With standardized workflows spanning immunization, cell fusion, screening, subcloning, expansion, and antibody purification, we deliver monoclonal antibodies with defined specificity, reproducibility, and complete documentation to support research, assay development, and early-stage therapeutic exploration.

Nebulabio Single B Cell Sorting Platform

Hybridoma technology immortalizes antigen-specific antibody secretion by fusing B cells from immunized animals with myeloma cells to create stable cell lines that continuously produce monoclonal antibodies. This platform is widely used for antibodies against conformational epitopes, cell-surface targets, and complex native antigens, and remains a robust route when direct immune selection in vivo is desired. Our end-to-end service covers immunization planning, fusion and selection, multi-round screening, monoclonality assurance, production scale-up, purification, and project-aligned analytical characterization.

☞Supported Options

◆ Host Species

• Mouse (BALB/c, C57BL/6, and project-configured strains)

• Rat (Sprague-Dawley, Wistar)

• Transgenic hosts (e.g., human Ig-expressing mice) when configured

• Species selection aligned with antigen characteristics and downstream objectives

◆ Immunization Strategies

• Recombinant proteins, peptides, conjugates, or cell-based immunogens

• Adjuvant selection configured to antigen class and response profile

• Immunization schedules aligned to titer development and specificity

• Pre-bleed and serum titer monitoring when configured

◆ Fusion and Screening Methods

• PEG-mediated fusion or electrofusion routes when configured

• HAT selection for hybridoma enrichment

• Primary screening by ELISA, flow-based binding formats, or microscopy-based binding formats when configured

• Isotype determination (IgG, IgM, IgA)

• Secondary validation for specificity and cross-reactivity

◆ Cell Culture and Cloning

• Hybridoma expansion in serum-containing or serum-free media when configured

• Limiting dilution subcloning for monoclonality assurance

• Cryopreservation and master cell bank generation

• Stability and productivity assessment across passages when configured

◆ Antibody Production and Purification

• Small-scale culture for screening and early characterization

• Scale-up in flasks or bioreactor formats when configured

• Purification by Protein A/G, antigen-specific affinity, or configured capture systems

• Buffer exchange, concentration adjustment, and sterile filtration

☞Principle of Hybridoma Technology

Hybridoma technology leverages in vivo immune selection to generate monoclonal antibodies with defined antigen specificity. Following immunization, antigen-primed B cells are harvested—commonly from spleen—and fused with immortal myeloma cells to create hybridomas that combine antibody secretion with continuous proliferation. After HAT selection enriches for fused cells, supernatants are screened to identify antigen-reactive clones. Positive clones are subcloned to ensure monoclonality, expanded, and used for antibody production. The resulting hybridoma lines provide a renewable source of monoclonal antibodies without requiring genetic engineering of antigen-binding regions.

☞Nebulabio Service Workflow

☞Featured Applications of Single B Cell Sorting

◆ Therapeutic Antibody Lead Generation

Hybridoma-derived antibodies provide in vivo-selected binders that can serve as starting points for early-stage therapeutic evaluation, particularly when native conformational recognition is required. Stable secreting lines support consistent supply during discovery-stage profiling.

◆ Diagnostic Reagent Development

Hybridoma technology enables generation of specific antibodies against biomarkers, pathogens, or custom antigens for assay development in plate-based immunoassays and cell-based binding formats. Clone stability supports continuity across assay optimization cycles.

◆ Research Reagent Production

Monoclonal antibodies from hybridomas remain standard tools for antigen detection and localization studies. A renewable cell line foundation supports reproducible experimental workflows and lot-to-lot continuity.

◆ Vaccine and Infectious Disease Research

Hybridoma campaigns against immunogens derived from pathogens support epitope mapping, neutralization studies, and establishment of reference reagents for immune monitoring assays.

◆ Oncology and Cell-surface Target Programs

Hybridomas generated against cell-surface receptors and tumor-associated antigens provide binders suitable for target validation and mechanistic studies in cancer biology contexts.

◆ Autoimmune Research and Autoantibody Isolation

In disease models, hybridoma workflows can isolate monoclonal autoantibodies that support mechanistic investigations and antigen identification in autoimmune settings.