Keeping research pure

The antibody market is a huge worldwide industry, but how do scientists know that what they are ordering is of the best possible quality? Dr Giancarlo Barone explains the current predicament for researchers and his attempts at resolving this issue.

With over two million antibodies for scientists to choose from, this particular market catering the research sector is both large and confusing. The current market is worth an estimated $2.6 billion per annum and growing at a rate of 5%.

For someone on the outside looking in, the protein binding market which serves the research sector is both successful and flourishing. While this may be the case for antibody suppliers, the positive facts above don’t truly reflect the frustrating problem for scientist’s past and present. Nothing displays this more clearly than a recent commentary in Nature¹ which highlighted a staggering waste on poor quality antibodies at 50%. While 50% of all antibodies purchased seems bad enough this does not reflect the true burden on the researchers grant. This is because scientists often spend significant research time and resources optimising conditions and validating recently purchased antibodies. Therefore, the true waste of research funds are no doubt considerably under reported. With this, let’s consider further the antibody market, its pitfalls and available resources that aid researchers in antibody purchases and consider how the scientific community can conjoin to build an antibody resource for the future.

[caption id="attachment_54961" align="alignnone" width="620"] How do scientists know the purity of the antibodies they are ordering?[/caption]

There are over 130 suppliers of antibodies, some are much better than others and we all have our favourites for particular reasons. At worst, suppliers offer scant information about how the antibody was raised or which, if any, application(s) the antibody has been tested in before being put in the shop window. Even the better suppliers fall short of the mark and really need to do more. Suppliers need to take more responsibility in thoroughly testing their product before sending it to market and stop being over-reliant on the end user to do this for them. I struggle to think of a similar scenario were the consumer readily spends £300, not knowing if the product will work and with the knowledge that they will probably spend time and money to find out. Maybe some of the problem here is that the end user is not spending their own money. If they were, consumer pressure would probably push suppliers to be more accepting of fewer profits over a better quality product.

For someone on the outside looking in, the protein binding market which serves the research sector is both successful and flourishing.

This leads me to highlight three particular areas which, if improved, would help scientists considerably in cutting antibody research costs.

Standardised data sheets The variability in both the content and quality of data sheets that accompany each antibody needs to be standardised. Each antibody should show general information relating to the antigen, batch number, catalogue number and clone identification. This holds particular importance when the same antibody clone is sold by several different suppliers under a different guise. Each antibody should be tested in at least one application, with the cell line, diluent, concentration, incubation period and actual experimental data provided for each application. This really should not be a problem given that each antibody is presumably tested by the supplier.

Antibody validation Antibody validation by knockout (KO) using siRNA or CRISPR is by far the most important point. If suppliers made such a practice standard then the so called antibody crisis would be pretty much resolved overnight. Instead, very few antibodies are validated before being sold by suppliers and as above it is left to the will of the end user to spend time and money to first validate their antibody. To be fair, it is worthy to note that a number of more conscientiousness antibody suppliers have recently agreed to put in place a scheme to do more to validate antibodies by KO. While I welcome this positive move by suppliers, it is early days and I am somewhat cautious in how far reaching this movement will be. Indeed, one supplier which has been at the forefront of this recent movement has more than 50,000-60,000 antibodies in their catalogue and have been running the validation by KO scheme for over 6 months but have only validated less than 400 antibodies from their entire catalogue.

This leads me to suggest that creative thinking is required to revolutionise the antibody market — one where both suppliers and consumers work together.

Validation by KO needs to be accompanied by experimental data sets for each application tested. This is important because not all antibodies work well in all applications. In addition, the whole process of validation will need repeating once a new antibody batch has been created. While, validation is both welcomed and needed, I fear that suppliers may only validate the most popular antibodies in their catalogue. This is a reasonable argument for the smaller, less financially resourced suppliers, as validating every antibody in their catalogue would no doubt be loss making. This leads me to suggest that creative thinking is required to revolutionise the antibody market — one where both suppliers and consumers work together. An example would be a scheme where suppliers offer free reagents such as an antibody and pool of siRNA for each gene of interest. In return for free reagents the scientist would be contractually obliged to produce quality data as per the supplier’s request. This way the supplier’s financial outlay and time constraints are greatly reduced while scientists receive free reagents and identify antibodies that they feel confident with and, more importantly, independent robust data sheets are produced for the supplier to host.

Batch to batch variation Antibody quality from one batch to the next can vary substantially. It’s incredibly frustrating to find an antibody that works well for several years to suddenly find the new batch no longer works. It is important for antibody suppliers to make sure new batch numbers are clearly illustrated on data sheets and accompanying experimental data is updated when a new batch of antibody is produced. This will be important if the antibody market commits to roll out the validation scheme throughout their entire catalogues.

There are number of free resources that aim to help scientists in their antibody purchases. Generally these can be broken into two categories, citation based and antibody review search engines.

Citation based search engines. The most notable website is CiteAb, but others exist. Simply type the antibody name into the search function and it compiles a list of research articles matched to the list 130 or so suppliers. This is a great resource for scientists and one which I highly commend. However, it has its limitations on several levels. Firstly, not all research articles provide sufficient antibody methodology and often don’t provide any validation by KO data. Finally, because of batch to batch variation, research articles that are a few years old no longer represent the original antibody cited in the article.

Antibody review search engines. Most notably, Antibody Adviser but there are a few others. These websites host antibody reviews and while they clearly set out to resolve an obvious problem, they don’t meet the necessary expectation. A review site of this nature needs to address the issues outlined above with more accuracy to earn the trust of scientists. A successful antibody review website should only host first class, independent and thorough reviews that surpass what antibody suppliers put on their data sheets.

At the time of writing I am about to embark on a campaign to raise funds to produce a free and independent review website. I think I have clearly set out what an antibody review website should provide to the scientific community. However, I strongly believe that a successful website needs to provide more and give something back to scientists in return for submitting comprehensive reviews. My vision is provide a user friendly website for the scientific community that combines a review website with a peer-to-peer support forum with endless useful research tools such as on-site Private Messaging, Protocols Deposit Box and invited guest Vloggers.

The dilemma is how to raise the necessary funds. My chosen route is crowdfunding and is being supported by FutSci who specialise in science related crowdfunding.

Author:

Dr Giancarlo Barone is a senior postdoctoral researcher with experience in DNA damage repair signalling and is currently working in the Genome Stability Group at the Department of Oncology & Metabolism, University of Sheffield Medical School.

References: 1 Andrew Bradbury & Andreas Pluckthunn, Feb 2015

Contact: If you wish to follow or support my antibody review campaign then visit www.antYbuddY.com or visit Twitter: @Anty_Buddy.