Single B Cell antibody discovery technology

Medicilon can provide Single B Cell antibody discovery service, which contains Single B Cell Sorting, Single B cell Ig Gene Transcription, Amplification, and Sequencing. The principle of single B cell antibody discovery technology is that each B cell contains only a pair of functional heavy chain and light chain and each B cell has the characteristic of producing only one specific antibody.

Monoclonal antibodies (mAbs) are important tools for many diagnostic applications and research. In addition, monoclonal antibodies have become an ideal therapeutic product for many diseases, and have been well used in the treatment of cancer, autoimmune diseases and infectious diseases. A number of techniques are used to generate monoclonal antibodies for research and therapeutic purposes. Single B cell screening is a newly developed technique for rapid preparation of mab in recent years. The principle is that each B cell only contains a pair of functional heavy and light chains, and each B cell only produces aspecific antibody characteristic, which can be directly amplified from a single B cell to obtain mab. This method has the advantages of fast speed, high throughput, and natural pairing of variable regions of antibody weight and light chains. It is one of the new and efficient methods for antibody discovery.

Process of Single B Cell Antibody Production
The Concept of Single B Cell Antibody discovery technology
The principle of single B cell antibody discovery technology is that each B cell contains only a pair of functional heavy chain and light chain and each B cell has the characteristic of producing only one specific antibody. Therefore, the antibody coding sequence can be amplified directly from a single B cell to obtain a monoclonal antibody.
Single B Cell Sorting
Depending on the research application, single B cells can be isolated randomly or antigen-selectively from peripheral blood or lymphoid tissues (bone marrow, spleen). For the isolation of B cells, current single B cell sorting technology include flow cytometry, magnetic bead cell sorting, micromanipulation, laser microdissection, and microfluidic sorting. Fluorescence activated Cell Sorting (FACS) is a mature and effective single cell sorting technology.
Single B cell Ig Gene Transcription, Amplification, and Sequencing
Construction of complementary DNA (cDNA) from a single B cell provides an efficient method for simultaneous analysis of expressed IgH and IgL genes. Typically, single B-cell cDNA synthesis is performed in equipment used for cell deposition and cell lysis (96-well plates, nanowell chips, etc.), which ensures easily handling of large numbers of samples and minimizes the risk of cross-contamination. Full-length Ig variable region gene transcripts were amplified by nested PCR, and RT-PCR products were used as samples for the first round of PCR for further reactions.
Regardless of the Ig variable region gene amplification strategy used, the transcriptional information of the Ig variable region genes encoding antibody-specific single B-cells was subsequently sequenced. Then using various databases such as NCBI's IgBLAST and IMGT, the rearranged V, D and J gene segments can be easily identified and analyzed for mutations, insertions and deletions.
Case Study: Nectin-4 Mouse Serum Titer
Case Study: Nectin-4 (4-1) Sorting Single B Cell
Case Study: Nectin-4 (4-1) Sequencing (Top20)
Case Study: Nectin-4 (4-6) Sorting Single B Cell
Case Study: Nectin-4 (4-6) Sequencing (Top20)

What is preclinical testing?

In the process of preclinical testing of a compound or biological agent into a drug, the compound involved must go through the testing phase. First, we need to identify potential targets that can treat the disease. Then, a variety of compounds or preparations are screened out. Any compound that has shown potential as a drug for the treatment of this disease needs to be tested for toxicity before clinical testing to reduce the possibility of injury. preclinical testing What is the basis of preclinical testing? According to US Food and Drug Administration (FDA) regulations, a series of tests are required before a new drug is approved for use. In the first stage, basic research determines a hypothetical target for the treatment of a certain disease, and then screens small molecules or biological compounds to discover any substance with the potential to treat the disease. Then, a preclinical research phase followed, before which, as described above, the potential toxicity of the compou

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Inventory of the three major in vitro pharmacokinetic research methods

The metabolic properties of a compound are an essential factor in whether or not it can be used as a drug in the clinical setting, so pharmacokinetic studies of newly synthesized compounds are required in drug development. In vitro incubation with liver microsomes, recombinant CYP450 enzyme lines, and in vitro incubation with hepatocytes are some of the more common in vitro drug metabolism methods. 1. In vitro incubation method with liver microsomes The metabolic stability and metabolic phenotypes of candidate compounds in different species of liver microsomes are good predictors of the metabolic properties of compounds in vivo. They are practical tools for evaluating candidate compounds in the pre-development phase of drug development. Liver microsomes include rat liver microsomes, human liver microsomes, canine liver microsomes, monkey liver microsomes, and mouse liver microsomes. In in vitro incubation of the liver, microsomes are the "gold standard" for in vitro d

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Novel Parkinson’s Therapies Possible with New Mouse Model

Parkinson's disease (PD) is a neurodegenerative disorder that is marked by the accumulation of the protein, α-synuclein (αS), into clumps known as Lewy bodies, which diminish neural health. Now, researchers from Brigham and Women's Hospital (BWH) report the development of a mouse model to induce PD-like αS aggregation, leading to resting tremor and abnormal movement control. The mouse responds to L-DOPA, similarly to patients with PD. The team's study (“Abrogating Native α-Synuclein Tetramers in Mice Causes a L-DOPA-Responsive Motor Syndrome Closely Resembling Parkinson’s Disease”) on the use of this transgenic mouse model appears in Neuron . “α-Synuclein (αS) regulates vesicle exocytosis but forms insoluble deposits in PD. Developing disease-modifying therapies requires animal models that reproduce cardinal features of PD. We recently described a previously unrecognized physiological form of αS, α-helical tetramers, and showed that familial PD-causing missense mutati

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