Antibody Discovery Using Single B Cell Screening Receptor Sequencing

The COVID-19 pandemic has made antibody discovery a major topic. There are many technologies that can produce antibodies. Hybridoma technology, which immortalizes antigen-specific B lymphocytes in order to create monoclonal antibodies, has been the standard for producing monoclonal antibodies for many decades.

Hybridomas can be cells that are formed from a combination of an antibody-producing B cell which is short-lived, and an immortal myeloma-cell. Every hybridoma is able to express a high amount of monoclonal antibodies (monoclonal antibodies). A stable hybridoma can be stored in cryopreservation to provide long-term monoclonal antibody production. There are many technical steps involved in creating hybridomas. These include antigen preparation, animal immunization and cell fusion. Sub-cloning and characterisation of specific antibodies is also required. Hybridomas are a great tool for discovering and producing antibodies for many applications. However, there are some drawbacks to the technology. There are several disadvantages to hybridoma technology, including contamination of hybridoma culture, limited range of useful antibodies generated due to losses during the fusion process and genetic drift over time that can lead to batch effects, cost, maintenance time and limitations for use in rats and mice. The identification and generation of high-quality hybridoma cells is a labor-intensive and slow-paced process that requires several months of effort from immunization through specific hybridoma identification.

Hybridoma technology’s limitations are overcome by newer methods of antibody discovery such as phage-display technologies. MedGenome, however, has standardized (High-throughput antibody detection using single-cell sequencing). This streamlines the process for antibody discovery using high throughput single-cell-cell receptor sequencing (scBCR–seq). It allows for precise paired full-length variables regions in a massively parallel manner. This method is able to quickly discover thousands of antibodies by using single-cell BCR sequencing. It also bypasses hybridoma based antibody discovery. When combined with high-quality antigen specific B-cell sorting, scBCR–seq can be used to quickly discover large and diverse panels of antigen-specific antibodies that are high in affinity. GenScript ProBio is superior to other antibody discovery methods because it examines each B-cell individually. GenScript ProBio gives users a prioritized list with candidate sequence panels. This includes immunizations of mice and rats with the antigen of their interest, as well as isolation of individual cells. The antibody-forming cells are then taken from the B-cells. Each B-cell is processed using the 10X Chromium platform to prepare single-cell libraries and sequence them for identification of paired heavy and light (Vh) chain antibodies.

GenScript ProBio offers key advantages over traditional hybridoma-based antibody discovery.

It does not depend on Single B Cell Screening and instead relies upon isolating the B-cells of immunized rats and mice from their spleens or lymph nodes.

It is cost-effective and time-efficient to obtain a panel paired prioritized antibody sequences for testing/screening within 12 weeks, as opposed to 6-8 months for hybridoma based antibody discovery.

Maximal antibody diversity is superior to hybridoma, which has a lower B-cell repertoire because of fusion loss.

scBCR–seq is not limited to humans and mice. MedGenome can provide custom species, including horse and rats.

Antibody Discovery by Beacon Single B Cell Screening

A popular microsystem-based screening technique that uses Single B Cell Screening or sort is being increasingly adopted in the field of antibody discovery. Berkeley Lights’ Beacon platform combines the most recent opt fluidics technology with high-throughput platforms to assay screen and integrate the screening, evaluation, and isolation of B cells on one chip.

One-cell Screening Method for Selection and Recovery of Antibodies

Antibodies are crucial tools of the adaptive immune system and have been used for innovative therapeutics such as cancer immunotherapies and vaccines against infectious diseases. Modern medicine has been unable to use antibodies due to many advances in antibody discovery.

Combinatorial display and hybridoma-fusion are the most common methods for antibody discovery. Vanderbilt University researchers have now suggested that single-cell transcriptomic methods could be an emerging method of antibody discovery.

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