Monoclonal antibody (mAb) as the pioneer of antibody drugs, more than 100 monoclonal antibodies have been approved for the treatment of cancer, infectious disease, own drug development market, and show their unique therapeutic value. Single B cell antibody technology, as a new generation of antibody development technology, can efficiently and quickly isolate antibodies from single B cells, which is a breakthrough in antibody production technology after hybridoma and phage display. Single B cell sorting technology can be used to antibody generation against multiple species such as rabbits, mice, camels, sheep, and chickens. Single B cell technology has outstanding advantages such as high specificity, high activity, and high affinity. Based on the Single B cell sorting platform, Alpha Lifetech has advantages in screening time and obtaining high-quality antibodies. It can provide antigen design, synthesis, and modification, animal immunity, single B cell enrichment screening, single cell sequencing, high throughput screening, expression, and purification, One stop technical services such as functional verification to meet the diverse needs of customers.

Select the appropriate antigen to the experimental animals (such as mice, rabbits, etc.) for immune stimulation, so that they can produce specific antibodies against the antigen. 

Animal immunzation is the initial stage of antibody generation. Animal immunzation is the use of the humoral immune response mechanism of the organism itself, through the continuous stimulation of antigens, animals produce a strong immune response, and then secrete specific antibodies against specific antigens.

B cells were isolated from peripheral blood, spleen or lymph nodes of immune animals. These B cells have been immunologically stimulated to produce antibodies against specific antigens.

By physical or chemical methods (such as density gradient centrifugation, magnetic bead separation, etc.) enrichment of B cells, removal of impurity cells, improve the purity of B cells and the efficiency of subsequent separation.

Fluorescence-Activated Cell Sorting(FACS), Magnetic Cell Separation(MACS), Microfluidic Cell Sorter(MCS) or other high-throughput screening techniques, combined with antigen-specific labeling, can be used to screen individual B cells from enriched B cells to produce targeted antibodies.

When a single B cell is isolated from a mixed B cell population, its antibody genes need to be sequenced by single cell sequencing technology and analyzed to obtain the DNA sequence information of heavy chain variable region (VH) and light chain variable region (VL). Single cell sequencing technology mainly includes three steps: single cell isolation, intracellular nucleic acid amplification and sequencing analysis. The principle of single cell sequencing is to amplify the trace whole genome DNA or RNA of isolated single cells to obtain a complete genome or transcriptome with high coverage for high-throughput sequencing. 

Through high-throughput screening, antibody gene sequences of thousands of B cells can be obtained at one time, greatly speeding up the speed of antibody discovery. These sequence information can be further used for antibody library construction and screening.

Fig.2 Schematic diagram of single B cell capture, library construction, high-throughput screening and sequencing. (Reference source: Massively parallel single-cell B-cell receptor sequencing enables rapid discovery of diverse antigen-reactive antibodies.）

(1)Recombinant protein: ≥2.5 mg, protein purity >85%, protein molecular weight greater than 10 kDa, soluble protein concentration between 0.5 and 5mg/mL. If antigen affinity purification is required, an additional 10 mg of recombinant protein is required.

(2)Polypeptides and small molecules: Bare peptide ≥10 mg, peptide purity >90%, no less than 10 AA, coupling KLH/BSA/OVA or other carrier proteins; Small molecules require structural evaluation in advance.