: What are the most important factors that a manufacturer should focus on when making an antibody?
CM: The keys to developing a great antibody product are in designing appropriate immunogens, screening for clones in relevant systems, and thorough validation of the clones and materials they produce. This is where the vast majority of effort must be spent. The failure to invest resources in this portion of product development is a key reason some companies fail to achieve a reputation for quality, despite having produced a large collection of antibodies.
: What aspect of your manufacturing process are you most proud of?
CM: The key to quality in manufacturing is to be able to reproducibly produce optimized material batch after batch. To do that successfully you should be able to measure and characterize the materials at all critical steps in the manufacturing process. We have stringent metrics at each of the many stages of our manufacturing process that allow us to ensure only high-quality materials pass from one stage to the next. Under our ISO13485 system we are continuously looking for improved processes that let us raise the bar on these metrics. After many years of manufacturing at high volumes, we have been able to incorporate a large number of process improvements which, in combination, have resulted in a very efficient process that yields exceptionally predictable antibodies and antibody conjugates.
: How does your company ensure the quality of the antibodies it produces?
CM: BioLegend manufactures antibody products at our main facility in San Diego, where we are certified to operate under ISO 13485:2003 standards. Our Quality Management System ensures traceability, reproducibility, and continuous improvements in manufacturing.
It is very important to us that every product that leaves our facility performs exactly as promised. This starts with validation of all new antibodies against endogenous targets in the applications that will be most relevant to customers. The confirmation of specificity and lack of off-target binding is confirmed with appropriate cellular controls, comparison with any pre-existing antibody clones, and appropriate performance in multiple relevant assays. Subsequent manufactured lots of any given antibody are characterized through several biochemical and functional QC tests.
: How do you ensure lot-to-lot consistency with your products ?
CM:
- By developing a high-quality clone, producing the antibody in a stable expression system (e.g. cultures using either hybridoma or recombinant cells), controlling production well, and carefully QC testing each lot, we can guarantee reproducible performance against endogenous targets.
- We have stringent quality systems that monitor each production lot and strive for continuous improvement over our already high standards.
- Each production lot is QC tested by a variety of biochemical assays, and in the most relevant functional assay for that antibody’s molecular target.
: What is your process for validating a given antibody for a specific application?
CM: Internally, we have stringent quality systems that monitor each production lot. During validation, we make use of any technology that allows us to develop the most relevant model systems and controls. These include knock-downs, knock-outs, blocking and competition assays, bench-marking to existing clones, and staining on endogenous targets. The data generated is made publicly available through our website. We have also begun citing contra-indicated applications when we have demonstrated an antibody is not suitable for a particular application.
: How does your company view the issue of antibody validation? Who is ultimately responsible for ensuring that scientific results are reproducible?
CM: It is our responsibility to validate the antibodies we commercialize. We must ensure that they recognize the correct target, do not recognize undesired targets, and we must provide as much context as we can regarding their optimal use. When we provide an antibody conjugate (e.g. fluor, enzyme, oligo) we must have proven that it will work readily in the application(s) we suggest. This is true for both the initial validation of a new antibody, and for each subsequent lot produced.
Ensuring scientific results are reproducible is the responsibility of both suppliers and end-users. Suppliers must ensure they are providing a dependable product. End-users must ensure they obtain their experimental materials from reliable suppliers, and then use such products in well-designed experiments that make appropriate use of the materials.
When scientists are validating an antibody for use in their experiments, they should:
- Always follow manufacturer’s recommendations
- Employ a relevant model system to test the antibody
- Use ample controls, both for the target (e.g. negative cell line) and for the reagent (e.g. isotype controls that are modified in the same fashion as your antibody)
- Compare to other similar antibodies if available
- If something looks amiss, contact the supplier’s Technical Service and share your data with their scientists.
: How do you incorporate customer feedback into your processes?
CM: We employ expert scientists to assist customers with trouble-shooting and experimental design. Providing direct access to these scientists is a key part of helping our clients achieve success.
Author Bio: Craig Monell, Ph.D., BioLegend’s Vice President of Business Operations, has 20+ years’ experience in the Biotechnology industry, including leadership roles in Marketing, Business Development, R&D, and Project Management at PharMingen/BD Biosciences, Bio-Carta, and Stratagene/Agilent Technologies.