: What are the most important factors that a manufacturer should focus on when making an antibody?
PS: When making a new antibody or manufacturing a new lot there are four important focus areas that need to be addressed by the manufacturer. These focus areas are high specificity, sensitivity, reproducibility, and application specificity.
: What aspect of your manufacturing process are you most proud of?
PS: We are proud of our comprehensive manufacturing process, which starts with antigen development and immunization and finishes with routine quality assurance testing. For new targets we design multiple immunogens, taking into consideration any post-translational modifications, structure, cross-reactivity, and species homology. In most cases we discuss target selection with leaders in key research areas and utilize their expertise to guide development and validation. This is followed by a multi-step, multi-application validation process. A very important step in this process is the technical review carried out by an independent team of scientists who critically examine each piece of data and only those antibodies that pass this stringent review are made available to our customers.
: How does your company ensure the quality of the antibodies it produces?
PS: Once the antibody is made it undergoes a multi-application validation process. We have an extensive tissue bank and blot library that allow us to test each antibody in a variety of applications to determine the precise specificity of the antibody. We not only concentrate on the detection of protein in a sample, but also its sub-cellular localization. In addition, we test our antibodies in several applications to assess specificity for the target and provide contextually relevant validation in conditions most likely used by researchers. Some of our antibodies also undergo enhanced validation, which involves the use of multiple antibodies for the same target, parallel knockout or knockdown techniques, and the use of orthogonal techniques whereby immunodetection results are compared with RNA-seq results to obtain an antibody-independent method for measuring the gene or protein of interest.
: How do you ensure lot-to-lot consistency with your products?
PS: We follow consistent protocols and keep retention lots for comparison. Our monoclonal antibody performance is highly consistent from lot to lot. However, with polyclonal antibodies some degree of variability is expected with different bleed times, but we ensure that they meet our specifications of selectivity, specificity, and sensitivity.
: What is your process for validating a given antibody for a specific application?
PS: Our antibody validation process is centered on the customer’s needs, which include demonstrating specificity to a target and in some cases sub-cellular location of the target in the context of different physiological states. Based on these criteria, the first validation may be Western blotting or ELISA where we use control cell lines. We may also use, in parallel, lysates from wild-type and knockout cells or cell lines that are typically null for the target protein. In some cases, cells may be treated with known activators or inhibitors of a particular target molecule, and lysate pre-tested for a specific post-translational modified state, such as phosphorylation, acetylation, or glycosylation. Further validation is conducted using immunohistochemistry, immunocytochemistry, or flow cytometry techniques to confirm specificity and cellular location of the target molecule.
: How does your company view the issue of antibody validation? Who is ultimately responsible for ensuring that scientific results are reproducible?
PS: Although there are no established and uniformly accepted requirements for antibody validation, it is of utmost importance that manufacturers provide details of their validation process and their data supporting the selectivity, sensitivity, and application specificity of their antibodies. Principal investigators should make sure that researchers working in their laboratory are sufficiently trained to handle these antibodies and conduct needed experiments following established protocols and obtain reproducible results. Attention should also be given to sample preparation to avoid loss of or degradation of proteins or their dephosphorylation that may be critical in some applications. In addition, experiments must include controls.
: How do you incorporate customer feedback into your processes?
PS: As a supplier, we are responsible for the quality and performance of our antibody in any specific application, and we are highly responsive to feedback on performance from customers. We place very high value on our customer comments and feedbacks. We offer live technical services in different time zones throughout the world to provide immediate support to resolve any issues that may arise. Our technical service team is highly trained and experienced. They keep accurate records of all performance-related issues, so trends can be identified and resolved quickly. Our sales team is also in contact with customers and seeks their input. Some of our new products are introduced based on recommendations by our customers and their needs. We work closely with researchers to improve immunogen design and antibody performance. We have a growing network of beta testers to continue validation. And each one of our antibodies is backed by our 100% Antibody Performance Guarantee.