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.

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.

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.

Researchers are central to the endeavors of maintaining research reproducibility. Here are some steps you can take today to improve reproducibility for the long term.

BioLegend embraces all methods of antibody production without losing focus on what matters most: the functional quality of our end product. We understand that the “reproducibility crisis” cannot be solved by improving just a single aspect of the antibody pipeline, which is why we are committed to having the highest quality standards for all stages of production.

Learn about the approaches we take to eliminate variation, and how our validation process ensures consistent quality of all our antibodies:

Downstream stages of production. A 2019 market survey found that many scientists believe recombinant antibodies have improved specificity relative to their hybridoma-derived counterparts. Controlling the protein sequence is one potential method to limiting variability, but other downstream procedures can affect all antibodies. This is why BioLegend performs rigorous structural and functional validation of each antibody batch as the final step, regardless of its source.

Structural validation ensures antibody purity

HPLC analysis of anti-mouse CD8a hybridoma antibody (clone 53-6.7) (left) and anti-mouse CD8a recombinant antibody (clone QA17A07) (right) post-SEC purification.

Functional validation ensures consistent lot-to-lot performance

Alexa Fluor® 647 anti-human Granzyme B Hybridoma Antibody

Two lot comparison of Alexa Fluor® 647 anti-human Granzyme B hybridoma antibody (clone GB11) staining on human peripheral blood lymphocytes. Cells were co-stained with FITC anti-human CD8 (clone RPA-T8).

PE anti-human CD56 (NCAM) Recombinant Antibody

Two lot comparison of PE anti-human CD56 (NCAM) recombinant antibody (clone QA17A16) staining on human peripheral blood lymphocytes (different blood donor than above). Cells were co-stained with APC anti-human CD16 (clone 3G8).

Learn more about our quality control process | Learn more about recombinant antibodies