Introduction
SH800S Cell SorterMonoclonal antibodies exhibit extensive application prospects and significant value in biomedical and clinical research fields. With the continuous development and innovation of biotechnology, the application areas of monoclonal antibodies will continue to expand and deepen.
Single B-Cell Antibody Preparation is a novel technology that harnesses flow cytometry (FACS) and other advanced analytical platforms for the rapid development of monoclonal antibodies. This process begins by utilizing fluorescence and basic light scattering capabilities of FACS to distinguish cell features and efficiently sort out the target single B cells. Once isolated, RNA is extracted from these B cells, then reverse-transcription to cDNA. PCR amplification is employed to detect distinct antibody gene sequences. These unique sequences are recombined to generate monoclonal antibody sequences, enabling antibody micro-expression, followed by purification. The resulting antibodies undergo comprehensive analysis, including ELISA and BLI assessment, to evaluate their activity and binding affinity. This rigorous screening process allows for the selection of optimal antibody candidates.
ACROBiosystems, as a global brand focusing on the biopharmaceutical development, is committed to providing diversified services that accelerate the development timeline and reduce research workload for our clients. Our single B-cell antibody development platform stands out from traditional antibody development methods, as it enables highly specific B-cell screening while preserving the natural pairing of the antibody heavy and light chains. This preservation is crucial for enhancing the affinity and functionality of the final antibody product, ultimately improving its effectiveness.
Core Advantages
Technical Roadmap
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 Services
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).
| Antibody Type | Service Period | Delivery Contents | Price |
|---|
Rabbit monoclonal antibody
Mouse monoclonal antibody | 12-16 weeks | 1. All antibody VH and VL sequences
2. Expressed plasmids or purified antibodies (customized to your needs)
3. Complete experiment report
4. Quality inspection report (COA)
| Customized based on your specific needs.
We will issue a tailored plan and pricing quotation for you.
Click to contact us |
Note: In addition to antibody development, we also offer downstream kit development services based on these antibodies
Case Studies
Single B-Cell Sorting after Immunization with Antigen A
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
Anti-A Antibody Activity Identification (BLI Platform)
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)
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