Single B Cell Solutions: Unlocking Rapid Antibody Discovery

Our technical roadmap for antibody development service outlines the strategic plan and steps required to achieve specific technical goals, from antigen preparation to the establishment of a PK assay. Key stages include:

1. Antigen Preparation:Antigens with a purity exceeding 90% are prepared, and 3 to 20 mg is required depending on the type of target antigen.

2. Animal Immunization and Potency Determination:Mice or rabbits are selected for immunization according to antibody species requirements.

3. Single B Cell Sorting.Following spleen extraction, single B cells are sorted using FACS.

4. Antibody Sequence Analysis and Expression Identification.Sequences of heavy and light chain variable regions of single B cells are obtained by single B cell gene acquisition assay. The antibody sequences are analyzed to yield multiple unique sequences as required. The obtained unique sequences are synthesized genetically and the plasmids are constructed for transient transfection into HEK293 cells for expression in well plates. The supernatant is collected for ELISA or BLI identification or blocking ELISA validation to screen positive clones.

5. Antibody Small-Scale Test, Purification and Identification. The initially screened positive clones undergo 10 mL small-scale expression and antibody purification. SDS-PAGE purity analysis, UV concentration measurement, and ELISA activity analysis are conducted for identification. A sandwich method is also used for PK (Pharmacokinetic) Assay development.

6. Antibody Production and PK Assay Establishment. The preferred paired antibody is selected for large-scale production. SDS-PAGE purity analysis, UV concentration measurement, and ELISA activity analysis are performed. A PK Assay is established, including standard curve generation, sensitivity assessment, matrix interference validation, etc., ensuring that the sensitivity meets the requirements for clinical PK analysis of drugs.

Our monoclonal antibody development services cover a wide range of targets, including target proteins, payloads, peptides, single-chain fragment variable regions (scFvs), and the drug itself (for the development of anti-idiotype antibodies).

Note:  In addition to antibody development, we also offer downstream kit development services based on these antibodies

Left: total lymphocytes sorted from spleen single cell suspension
Middle: sorting of total B lymphocytes from total lymphocytes
Right: sorting of target-positive single B cells from total B lymphocytes

A preliminarily screening of several candidate anti-A antibodies was conducted, and their activity was determined by the BLI platform, which successfully identify the target antibodies among the candidates. (Only a portion of the data is being presented here)