BioAccord LC-MS: Making Mass Spectrometry Available to the Masses

Earlier in 2019, Waters Corporation announced their BioAccord LC-MS System for biopharmaceutical analysis. The BioAccord System is a self-calibrating, self-optimizing, self-sufficient tool that promises its users simplified high performance liquid chromatography-mass spectrometry (LC-MS) analysis and high quality data. It is optimized for "unparalleled robustness and reproducibility", offering its users consistent results "day in, day out". We spoke with John Gebler, PhD, Director of Strategic Business Development at Waters Corporation to learn more about the BioAccord System and its applications. 

Molly Campbell (MC): Could you please explain why LC-MS is so suited for the analysis of biopharmaceuticals?

John Gebler (JG): I'm going to draw some comparisons between traditional small molecules and large biotherapeutic molecules. Small molecules are well defined as far as their structure is concerned, their molecular composition and even their shape. There are a number of technologies out there that can verify that structure in very high purity, and one who takes traditional pharmaceuticals would expect a drug to be 99% or greater purity. Companies do that, regulatory agencies expect that, and so you can define that.

When it comes to biotherapeutics, the drugs tend to be heterogenous and that naturally happens because most of these drugs are produced by cells, which have a lot of processes going on. For example, antibodies are the most popular biotherapeutics on the market today, and these are post translationally modified, meaning there isn't single defined structures for those type of molecules. Biotherapeutics also tend to be very large when compared to small molecules, and instead of talking about hundreds of Daltons, we must talk about hundreds of thousands of Daltons in size. The structures of biotherapeutics are therefore extraordinarily complex and require technology that can interrogate that complexity.

LC-MS has emerged as one of the best technologies to do that and allows us to look at structural details from a top-level view, a middle view and a very granular view. We can take the proteins apart by breaking them into pieces, look at those pieces and stitch that information back together to be able to tell what qualities the drug has and what's going on with it. There really is no other technology that can do that right now, and the beauty of mapping back is that when you get a map of something it is directly measuring either that molecule or a part of the molecule that you might have broken off, such as a digestion and so forth. So, it doesn't lie as far as what data is there.

You can further interrogate the data by manipulating the molecule and fragmenting it and looking at it in a variety of ways, which gives tremendous insight to the molecule itself or the protein itself. This is very accurate today. When it comes to analysis of biotherapeutics, LC-MS is probably the most powerful technology out there and is widely adopted in the industry.

MC: What are some of the challenges that scientists face when using LC-MS in biotherapeutics?

JG: One of the things that I like to point out is that any company that wants to be competitive in biotherapeutics is going to have to use this technology. A lot of the big pharmaceutical companies set the bar a long time ago and they were some of the first users of LC-MS in the field and for obvious reasons; it was clearly the only way they could look at these molecules in a meaningful way. However, the challenge is that the LC-MS systems are complex and not necessarily easy to use. They require a level of skill and capabilities to interpret the data correctly. I've talked to scientists across the world a lot through my career at Waters, and one point I'd make to them is that experience means everything. The more experience you have with the technology the more successful you're going to be with it which is why you tend to see that samples for LC-MS analysis are sent by various departments to centralized laboratories where the MS expertise resides. Now, that creates a burden for companies in that it requires them to find people who are experts in this field, or they must be willing to train people to be experts in this field. It is not trivial to do that. 

So, there's common knowledge in the industry today that one must be an expert to run this system, and here we're really talking about the MS end of things. LC has been used in the industry for decades and so there's a lot of skill in how to run a HPLC system, or a UPLC system, out there. But, when it comes to MS there's a lot of, what I like to call "art" in addition to the science of it.

Biopharma is one of the fastest growing segments in pharmaceuticals today worldwide. Before establishing themselves in any given country, companies definitely want to be able to tap into a pool of scientists with the appropriate level of LC-MS skills, whether that be the United States which wants to continue its strong path in innovation, or if you're in India where you want to have a strong presence in biosimilars, which are the generic versions of biopharma, you're going to have to adopt LC-MS. The experience in MS is quite finite throughout the world and so this becomes a challenge. If companies could deploy LC-MS more easily throughout their organization, they would probably adopt it in areas that they're currently not using it in that would give them useful information. It would allow them to make their drugs safer, faster, protect their position more, be in a position to troubleshoot more quickly and simply just be more efficient.

One of the other things that we're seeing happening a lot in the pharma industry is companies looking more seriously at how they could deploy LC-MS into all kinds of their processes and drug development stages in biopharma because it allows efficiency on their part, it gives them more useful information. A term, multi-attribute monitoring (MAM) has been coined where the goal is to put LC-MS into quality control (QC) labs to replace a number of conventional assays currently being used. This is more of a desire and wish at this stage, but companies do have very active programs in this area right now.

To accomplish this, one needs LC-MS systems that are robust, reproducible and easy to operate. LC-MS also generates mountains of data and so we need methods to make useful information out of the data that will allow companies to make meaningful decisions. 

MC: Do you believe that the current challenges and limitations of LC-MS within biopharmaceuticals has impacted its wider adoption?

JG: Absolutely. I actually lived in India for a year and return there often, and when I talk about deployability expertise, this was a country that just didn't have it at all but knew they needed to adopt it.

As I said, the number of people who are experts in this area is just simply finite. I told graduates that if they were to become an expert in this area in India that they were going to get instant employment. It would be useful for academics to really understand that they're going to make their students instantly employable if they have this skill because pharma wants it so badly.

For us at Waters, we look at it as an opportunity - can we create systems that can be deployable and are easier to use? Whether you're in discovery or development, late-stage development, or process development (or ultimately in quality control (QC)), can we create a system that has robustness, reproducibility and is easy to use, to be able to allow companies to deploy it into multiple areas?

In general,  we as consumers have high expectations for the technologies we use every day.  You can see that in mobile phones and tablets, or even simple things like an automobile. We expect a certain amount of intuitiveness and intelligence in these kinds of technologies, and so the generation that is rising right now expect that across the board. It's not to say that we're trying to un-skill people or suggest people aren't capable of doing this themselves, but I think the reality is that we use all these tools to get the information that we need. 


The BioAccord System pairs the  ACQUITY UPLC I-Class Plus with the newly-developed ACQUITY RDa Detector featuring SmartMS and is powered by UNIFI, Waters’ compliance-ready LC-MS informatics platform. Credit: Waters Corporation.

MC: How does the Waters BioAccord LC-MS system help scientists overcome some of the challenges you've discussed?

JG: One of the things that we did, and we started this a number of years ago, is interviewing scientists. We asked them to talk to us about their challenges in a very meaningful way, and we conducted a large number of interviews across the company. We started seeing a very common pattern emerge – a desire for robustness, reproducibility and ease of use. People also always tell us they want better informatics systems too.

So, there are multiple items that one has to address with a new LC-MS system: you have to generate high quality data and then you have to be able to process that high-quality data in a way that's fast and accurate. We recognized at Waters that this was an opportunity to create a new LC-MS system. This system could be bought by a person, company or scientist and it would be delivered to them as a system. It has been tested as a system. It has been crafted to work together in absolute concert. It is our BioAccord system. The system consists of our ACQUITY I-Class UPLC separations system, various chemistries for intact and sub-unit mass analysis as well as released-glycan analysis, and the heart of the system, which is the ACQUITY RDA time-of-flight mass detector for accurate mass analysis of peptides, proteins and glycans.

With the system we made a new time applied instrument that we call SMART-MS™ technology that is built in. Previously when you wanted to start an experiment, it would take many clicks in the software and from many different pages to set the experiment up. With the BioAccord system, everything’s presented to the operator on a single set-up page, and the choices can be made very simply as the page is optimized. For example, when you click on peptides, the system calibrates itself, goes through a diagnostic check and let’s you know when it’s ready to go.

Not surprisingly, some scientists were nervous about that. Researchers that we spoke to were used to poking around in the software and with the instrument and thinking "well I can really get this to sych", but when we did the beta testing of the SMART-MS their feedback was, "oh this thing works really well and it's easy to use". So, we knew we had basically nailed down the ease of use requirement. The new TOF MS is interfaced to the ACQUITY I-Class UPLC system known for its extraordinary reproducibility.

Then we went an additional step, at how we test our LC/MS systems. When you're in biopharma you're looking at an antibody that is 150KDa and you're looking at heterogeneity, and when you're looking at peptides you're looking at something that's very complex.

The opportunity/ challenge for us was to create samples that we could test the system with that actually reflect what people will actually do with it.  We locked the NIST antibody, which comes from the US government, recognized as a standard throughout the world, and we worked on obtaining that sample and packaging it up. We also created different versions of it for sub-unit and peptide digest versions of the test sample. We use those samples to actually test the system by LC- MS, and we had very strict criteria for the system to pass the test.

Before a BioAccord System leaves the factory for a customer, we test the complete system with the appropriate test sample to make sure it meets strict performance requirements. When it does, we put the gold star on it, and ship it to the customer in a single shipping container so that when the engineer installs it for the customer, the same sample is run on the system to ensure that nothing has happened during shipping, and again demonstrate to those who have made an investment in it that it actually meets those strict requirements in their lab.

We learned a lot in this process. We learned about how we had to refine our system integration testing  so that we maximise the speed of installing the system and then be able to demonstrate "here it is and it's been tested with something that's very real." It's created an interesting opportunity for us in that when we now demonstrate to potential customers and users, we can show them that this system is tested with the sample that they’re going to use. We find that people really appreciate that a lot and to our knowledge we're the only company that does this system integration testing. It's an expense that we have to absorb, but we believe that it has tremendous opportunity for us to really demonstrate to people that we are listening to them and we know that when we say this is a biopharma system, it is actually a biopharma system. 

We have additional things that we're going to build on to this. I mentioned a lot about robustness and reproducibility and ease of use, so we're also looking at our data processing software and how these applications are going to take better advantage of the high quality data. We've also paid attention to the informatics request because we believe that BioAccord is a very good system for collecting and recording data. We have our UNIFI informatics solution which is a compliance LC-MS solution, which is key if a laboratory wants to move the technology into late-stage development and eventually into QA/QC, right now, which has very good apps, but we're going to improve it even more by focusing on the needs of scientsits and the concept of deployability.

If you think about deployability, to give you a good example, lots of companies focus on optimization of their columns. So, right now that's done primarily by biologists and they tend to look at tighter in binding of the antibody, but they don't look at the drug product itself and say, "Is there anything I should be worried about?" Then a little later in the development process they realise "oh, oh, something's wrong with my glycan pattern", and if they could deploy MS sooner in their processes to be able to look at that, it would give them the information that they need to make better decisions. There's nothing worse for a bio-similar company than to realise six months down the road, that their bio-similar is not similar. There has always been hesitancy as to how you deploy sophisticated LC-MS technology in to that arena to prevent such mistakes, and here it is: the BioAccord system. Many people who have seen our system, whether that be through our virtual information that we have on the website or at scientific meetings, look at the data and say "oh, I actually might be able to do that," and its created a paradigm shift which makes us pleased and proud that we have made MS available to the non-experts.

Overall we’re very excited about the BioAccord System because it represents a new direction for Waters where we are taking the power of MS and putting it into the hands of non-experts, the people running the samples so they can take advantage of the benefits of the technology. 

MC: Is there anything else you'd like to add around system integration testing and how it may help broader adoption?

JG: Folks are going to see that we've really made an effort on how we test the system based on the samples they have available to them. When they worry about a performance, it would be easy for us to say well take sample number X, run it, and tell us what the result is. BioAccord itself is highly diagnostic and has built in intelligence to do that itself, so it's going to tell you where something may be wrong, and we are continuing to evolve that intelligence. I think that's where customers will really begin to recognize the value to them, in just having access to reference standards and also knowing how the system was tested and being able to say this is how I plan to use it and Waters is now supplying these things for me to be able to do that. High-quality standards are a big deal and are highly diagnostic to how well a system is behaving. We actually embarked on this path a long time ago because we found that as we tried to acquire these reference standards from various suppliers, that they were actually of a horrible quality. 

MC: In what way and what has changed?

JG: There weren't QC methods put in place and I'd say that manufacturing wasn't meant to be reproducible. There were different reasons why these kinds of things were being made and trying to solve different kinds of problems. For example, if you want to make a peptide digest from an intact antibody, one would just digest it with tripsen and let it sit around overnight and hope for the best. That's not very reproducible. Understanding the variabilities that control the reactions and making them so they're highly reproducible is what scientists need. At Waters we have focused on developing a digest standard that is consistent and that will ensure confidence and high quality results.

It is also important to look at how you package the material up, how to store it and maintain its stability. We are a global company and so also needed to ensure that we could ship it anywhere in the world. There are factors to consider. First, there's temperature: heating and cooling and those kinds of things are important to consider, and it took a lot of work, but in the end we've become very reliant on having these materials. They really help us create systems for our customers and for the biopharma and biotech world out there that they can feel very confident about.

John Gebler was speaking with Molly Campbell, Science Writer for Technology Networks.