Do antibodies have to be hit and miss?

From geneticist to frontline warrior in the antibody validation crisis – Dr Giancarlo Barone tells us how he is winning over the huge worldwide industry of antibody producers as he launches his review site antYbuddY.com

My journey began a few years ago whilst working on uncharacterised genes that play roles in Genome stability. The bulk of our work was validating our initial screen, deconvoluting siRNA pools into 4 different siRNA for each gene.

An important part of this work was using antibodies in Western blot (WB), immunofluorescence microscopy (IF) and immunoprecipitation (IP) to show the successful knockdown by siRNA and to confirm protein interactions identified in mass spec data. This is where I and other lab members ran into problems. More often than not we could not validate knockdown by WB or IF using commercial antibodies. In my case, I purchased 4-5 different antibodies for my gene of interest and only one worked in WB. The initial antibody costs alone was close to £1.2K with additional costs and several months wasted trying to validate knockdown by WB and IF. In all cases, we used RT-PCR as an alternative route to show successful knockdown.

Finding the right antibody is somewhat hit and miss, even when working on genes that have been characterised and I am confident that most scientists around the globe would agree with my statement. However, the fact that our lab was working on an uncharacterised gene of interest using commercial antibodies exacerbated the situation – these antibodies had clearly been through little or no validation process. Speaking from experience, I would guess there is a correlation between antibody success and gene popularity – if your gene of interest is already very well characterised then you’re one of the lucky ones.

These events got me thinking about creating an online tool where scientists could share their antibody experiences in the form of independent antibody reviews. I started the process by working with specialist science crowdfunding platform Futsci. Being a working scientist and a father of four young boys, this was not easy to juggle. I spent all of my free time, with late nights, early mornings and weekends sharing my crowdfund campaign on social media platforms which generated a lot of interest from the science community.

After 3 months of contemplating where to take this next, I decided to fund the antibody review platform – I had been campaigning for over a year by this point. I knew this was going to be a big commitment given how much effort my crowdfund had taken. It started with seeking the right website developer that would understand my idea but also work within my budget. I knew this would not easy, as building a website from scratch with the specs I wanted could easily cost upwards of £20,000. After many Skype meetings, emails and a well-considered plan in place my developer Alen Piljic began building antYbuddY at the end of January.

While all this was going on I was also trying to engage with antibody companies to present to them my vision and to assure them that antYbuddY.com is a worthwhile initiative. At the centre of all this was my idea of creating what I call sponsored ‘rewards’. I believe having rewards is important, the reviewer writes an antibody product review and is rewarded with an antibody discount, Amazon gift card or quirky gift. Trying to find contact details for directors, and global marketing managers of very large companies was a learning curve but getting them to engage is the really tough part. Without going into too much detail, persistence seemingly sometimes pay off. After a lengthy process, several companies – Proteintech, Novus Biologicals and Everest Biotech – now sponsor rewards on the antYbuddY platform.

In addition, several other leading antibody companies are looking to sponsor antibody reviews and I will continue to seek more sponsors to enrich the variety and scope of reviews. You may question what my site sponsors aim to get out of all this. The reason, is a sincere one. All our antYbuddY review sponsors clearly acknowledge the current issue with antibody validation but more importantly care enough to want to explore every avenue to remedy the problem and seek a close relationship with their custom base.

With antYbuddY built and several sponsors on-board, antYbuddY.com is now live. This is where the hard work begins. While continuing to juggle my job as a postdoc and family life I need to focus on marketing the antYbuddY platform to reach the worldwide community antYbuddY was built to serve.

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.