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New Antibody Development

BioLegend spends a considerable amount of effort in creating new antibodies for research. The majority of these new antibody products are monoclonal antibodies (mAb) produced from hybridomas. Clones of these hybridomas are carefully selected based on a number of criteria including robust growth and efficient production of a single clone of antibody that is specific to the intended target. The best clones move on to applications testing.

As manufacturers are starting to produce recombinant antibodies, these would need to be validated and addressed in the same ways as hybridoma-derived antibodies. A mAb produced from an engineered DNA source is not inherently more specific than one produced from a traditional hybridoma. However, genetic modifications to the overall structure could potentially convey improved performance in assays.

Applications Testing

All newly developed clones at BioLegend undergo validation testing for multiple applications. This serves as a cross-check for specificity and provides clarity for research uses. Typically, antibodies are tested by two or more of the below methods.

As an example, our newly developed clone, 13A3-1, for phosphorylated STAT3 (Tyr705) demonstrated excellent performance in flow cytometry, western blot, and chromatin immunoprecipitation. Thus, the clone cross-validates itself, by demonstrating functionality across orthogonal testing methods. Additionally, the biological induction of the phosphorylated state using IL-6 further validates the specificity of the antibody.

Clone 13A3-1 Application Validation

Clone 13A3-1 Application Validation

Gene Knockdown Testing

Knockout or knockdown of gene expression, such as with siRNA, is also an excellent tool for target validation. Our SIRT5 antibody, clone O91G9 was verified by western blot using HeLa cells treated with SIRT5 targeting siRNA. Lane 1 indicates untreated HeLa cells, lane 2 contains scrambled siRNA control treated cells, and lane 3 contains SIRT5-specific siRNA treated cells.

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BioLegend antibodies undergo an extensive series of testing to ensure quality at every step in the manufacturing process, as well as maintaining quality after the sale.



Maintaining lot-to-lot consistency is vital for reproducibility. It simply is not enough for antibody manufacturers to validate antibodies just once. Pass/fail specification requirements are essential for quality control testing of every lot of product. BioLegend maintains records for all lot-specific testing, so inquire with our Technical Support if you would like to see the data. 

An ISO 13485:2016 certified quality management system, which meets the requirements for medical device manufacturers, means that processes are well-defined, adhered to, and undergoing constant improvements. It also ensures that there is a robust system for customer notifications in case of non-conforming products. BioLegend’s 100% satisfaction guarantee provides peace of mind for researchers, so that they can be confident in the results every time.