: Can you share your perspective on the reproducibility issue associated specifically with the use of antibodies?
RP: Despite negative reports regarding scientific reproducibility, I believe there are many good commercial antibodies in the marketplace that are being used to make new important discoveries in biomedical research every day. However, there are also bad antibodies and companies that commercialize antibodies without proper validation. In our experience, this is the result of companies selling antibodies they did not develop or taking shortcuts to diminish the costs associated with solid validation and quality control, which can be expensive. A responsible antibody vendor should provide clear data to support claims made regarding the performance, limitations, sensitivity and specificity of any product they sell and stand behind their reagents with excellent technical support.
Beyond antibody suppliers, there are other stakeholders that may contribute to the reproducibility problem. First are antibody users who may not be familiar with antibody-based methods and/or do not follow the recommended protocols. Antibodies can be tricky to use, particularly for some complex applications. Deviation from the recommended protocols used to develop and validate the antibody by a vendor can lead to poor results even with a highly specific and sensitive antibody. Secondly, we believe journals publishing scientific papers need to do a better job compelling authors to provide all the necessary information about the antibodies and protocols used in the study. Insufficient information can make a great paper virtually irreproducible.
: Can you tell us what your company is doing on a routine basis to make sure you are testing and validating the antibodies you manufacture or obtain from other sources?
RP: CST is not just an antibody product company. Since our foundation, we have also done important basic and applied cancer research. We invest heavily in new technology development and our internal research programs. Like our customers, we also publish our research, so we understand the cost of failed experiments and the importance of researchers’ time. For these reasons, we only commercialize antibodies that were made and tested by us and for which we can provide the protocols and technical support to make sure they work in the hands of our customers the first time and every time. We place very strong emphasis on application-specific validation, quality control and technical support. For CST, product quality is simply a matter of straight business ethics. Our antibody development process has a high attrition rate. We would never sell an antibody that doesn’t work or is of poor quality. Thus we don’t have a very large catalog, but what we offer is likely the best quality in the market.
: Do you have any advice, recommendations and/or best practices scientists should follow when selecting and validating an antibody?
RP: My recommendations are: first, when purchasing an antibody always go to the highest quality providers. If you are new to the field, ask your colleagues. Second, look for validation data supporting the use of an antibody in the specific application of interest (Western blotting, IHC, etc) provided by the vendor. Look at the data critically as if you were reviewing a manuscript. Third, look for data supporting the use of the antibody in the desired application within the peer-reviewed scientific literature. Publications are not perfect, but I believe well-published antibodies are a good place to start. Fourth, try to strictly follow the vendor’s protocol, if available, before you introduce experimental variations. Finally, if you end up buying an antibody for which no data is provided and there are no published citations available, beware. You should first determine its specificity using as many application–specific positive and negative controls as possible.
: What are your thoughts on the current efforts that are underway to tackle the reproducibility problem? Do you think they are realistic and implementable?
RP: We believe the current discussions around the reproducibility problem, for example the Asilomar conference sponsored by GBSI, have the potential to be very useful for the community. We are actively engaged in the discussions and in efforts to design achievable validation standards. There is currently a significant push to design a scoring system to qualify antibodies in the marketplace. It is an interesting idea, but I have serious doubts about its feasibility, impartiality, cost and potential unintended consequences. There hasn’t been a serious discussion about this, in my opinion. Other ideas practiced by other industries such as some type of quality certification have not been discussed much, but I feel this is one potential alternative to scoring individual commercial and academic antibodies.
No validation guidelines will be complete without engaging all stakeholders to tackle the different factors contributing to the reproducibility problem, not just the antibody providers. Encouraging the training of young scientists in the use of antibodies is crucial—the responsibility of this task can fall on both academic institutions and vendors by ensuring that students have access to resources and information. Enforcing journal publication standards that require authors to clearly identify the source, catalog and lot number of any antibodies used and thoroughly describe methods is just as important and potentially easier to accomplish in the short-term.
: What else is your company doing to address reproducibility?
RP: The most important thing CST can do is to continue to work hard to maintain our industry leadership and efforts in antibody validation and overall quality. Our customers know our quality reputation, and we will maintain that. We are constantly improving our methods and technologies to do things better. Whether it’s the use of CRISPR technology or mass spectrometry-based methods, we continue to expand our capabilities to deliver to our customers the best products we possibly can. We will continue to help educate our customers by making available protocols, tutorials and scientific material on our website and by different outreach campaigns to help scientists succeed in their projects.
Actively participating in the debate and contributing to solve the reproducibility problem as part of the scientific community is very important. We will continue to do so. But we also hope to help create an industry forum or consortium where the reproducibility problems and solutions can be debated to help elevate the quality of our industry as a whole. As these changes take hold, the research community that we are a part of will benefit and we’ll begin to see new scientific discoveries made more reliably and in less time.
Author Bio: Roberto Polakiewicz, Ph.D., CST’s Chief Scientific Officer, has been responsible for Product and Technology innovation since 2005. His leadership has created the company's most valuable products and intellectual property. Dr. Polakiewicz has driven the successful implementation of four generations of novel antibody platform technologies critical for CST’s ongoing success. Prior to joining CST, Dr. Polakiewicz received his B.S. in Chemistry and Ph.D. in Molecular Biology from the Hebrew University of Jerusalem and completed postdoctoral studies at the Whitehead Institute for Biomedical Research at M.I.T. Dr. Polakiewicz has published approximately 66 peer-reviewed publications, and is a named inventor on numerous patents and patent applications.