You want to develop a monoclonal antibody conserving its native properties with maximized developability? With our single B-cell screening service, get a highly affine antibody preserving a natural VH and VL pairing and a maximum B-cell diversity. We guarantee you receive minimum 3 antibody sequences within maximum 2 months from animal immunization until antibody screening thanks to our high throughput screening platform.

Workflow of Single B Cell Antibody screening

IMMUNIZATION
  • Antigen checking by SDS-PAGE
    (Antigen provided by You or designed by us)
  • Rabbit immunization
    4-6 rounds of injections using an optimized protocolImmunization is performed on Rabbits

 

6-8 weeks

SINGLE B-CELL
SORTING & SCREENING
  • Isolation of PBMCs and spleen lymphocytes
  • Fluorescence-Activated B-Cell Sorting (FACS) antigen-specific single B-cells sorted (1cell/well)
  • B-cell culture and supernatant screening against antigen in ELISA

 

2-3 weeks

Want to test additional clones?

Additional sequences delivery

RECOMBINANT RABBIT ANTIBODY
(IgG) PRODUCTION & SCREENING
  • Best positive clones sequencing
  • Transient expression into high performance proprietary XtenCHO™
  • B-cell culture and supernatant screening against antigen in ELISA
Want additional ELISA?

ELISA for positive/negative screening
(Against one specific peptide/protein/small
molecule- cross reactivity profiling)

Competitive ELISA
(To identify clones with blocking/
neutralizing activity)

Advantages of Single B cell sequencing

Single B-cell sequencing is an efficient monoclonal antibody development and screening strategy which principle is based on the amplification of genes encoding the VH and VL region from B-cells directly. This technology presents several advantages making it a very useful approach for antibody biology understanding and for antibody use in clinical essays. Among these advantages, single B-cell sorting is characterized by:

  • High throughput B-cell sorting – Single B-cell sequencing integrates high throughput platforms for antibody selection which allows to perform different screenings in a highly efficient manner
  • Native monoclonal antibody preservation – The main advantage of single B-cell sorting technology is that it preserves the native VH and VL pairing, two domains of monoclonal antibodies critical for antigen recognition and binding. Thus, natural cognate pairing is maintained and development of antibodies with high affinity, stability and specificity is guaranteed.
  • Rare antibodies identification – Some antibodies with relevant therapeutic properties are hard to identify. Thanks to the single B-cell screening approach, some rare antibodies are identified directly from B-cells and this limits the risks of unnatural pairing resulting from in vitro screening. Moreover, this approach is ideal for discovery of rare antibodies against challenging targets such as conformational epitopes.
  • Antibody diversity maximization – Thanks to the highly sophisticated screening method, single-B cells are screened individually leading to a various repertoire of monoclonal antibodies which increases the B cell diversity.
  • Higher efficiency and expedited timelines – Single B-cell screening is a very efficient technology characterized by an unprecedented single-cell-resolution and an exceptional rapid timeline saving months of work as a result of avoiding the cell fusion (hybridoma) and library construction (phage display) steps.
Pro Cons
Hybridoma
  • High antibody affinity
  • Preservation of natural VH-VL pairing (positively impacts developability)
  • Cheap
  • High loss of diversity due to low efficiency of cell fusion
  • Limited to rodents as myeloma cells not efficient in other species
  • Humanization needed
  • Not adapted to antigens with low immunogenicity and/or toxicity
  • Animal use
Phage Display
  • No species restriction
  • No need for humanization thanks to fully human libraries & humanized mice
  • Maximized diversity due to random VH-VL pairing
  • High-throughput screening allowing identification of multiple binders
  • Adapted to antigens with low immunogenicity and/or toxicity
  • Very fast
  • No animal use thanks to premade libraries
  • Non-natural VH-VL pairing can negatively impact developability
  • Potential bias due to phage particle
  • Low/medium antibody affinity due to lack of immunizations (naïve libraries)
  • High affinity antibodies expensive & long to develop as require custom immune library construction
Single B-Cell Sorting
  • No species restriction
  • High antibody affinity
  • Preservation of natural VH-VL pairing
  • High-throughput screening
  • Best for discovery of rare antibodies against challenging targets (e.g. conformational epitopes) with maximized developability
  • Not adapted to antigens with low immunogenicity and/or toxicit
  • Animal use