2012 PEGS

This year marked the 8th Annual Protein Engineering Group Summit (PEGS) held at the historic Boston Park Plaza Hotel and Towers in Boston, Massachusetts which included over 220 scientific presentations held in 4 concurrent streams: Discovery, Expression, Analytical and Antibodies. Clearly the focus of the conference centered on biologics with streams such as Phage and Yeast Display, Engineering Antibodies, Characterization of Biotherapeutics and Antibodies for Cancer Therapy defining the majority of the sessions. The conference had ~1300 attendees visiting over a five day period and more than 100 poster presentations covering a wide range of applications from target identification to lead optimization to clinical data. More than 60 vendors displayed the latest technology and tools to aid in all aspects of protein engineering and processing.

One notable return to the forefront of biological agents was evident with a stream dedicated to Antibody-Drug Conjugates (ADCs). Many will recall long ago, about 15 years back, when many considered the development of ADCs as potential “magic bullets” that would be developed as highly specific agents targeted against cancer and other such diseases. ADCs are antibody or antibody derived scaffolds linked to a cytotoxic payload that will be highly specific for a therapeutic target. The cytotoxic payload will only be released once the target has been reached thus potentially avoiding adverse affects that accompany treatment with many therapeutic agents currently on the market. New molecular scaffolds, linker chemistries and toxic payloads are providing breakthrough technologies that researchers anticipate will finally help bring novel ADC drugs to the market, as evidenced by the recent approval of brentuximab vedotin (marketed as Adcetris) in August, 2011. Coupled with new insights into mechanisms of immunogenicity, novel assay technologies and improvements in bioprocessing the field of biological therapeutics appears to be gaining momentum with hundreds of clinical trials currently underway.

Presentations were held in several locations within the hotel covering a wide range of topics from overcoming challenges in protein expression to bispecific antibody optimization to up-to-date clinical trial data.

Spring was evident with the greening of Boston Common.  Unfortunately it had to be viewed under cloudy or rainy skies for most of the event.

Kilimanjaro and Hypoxia

Kilimanjaro rises to 5,895 meters (19,340 ft) where the amount of oxygen available for breathing is about half that relative to the surrounding plains of Tanzania.  At altitude the air we breathe becomes progressively thinner.  About half the atmosphere of the earth is within 18,000 ft of the surface - the remaining half is spread in a ever diminishing concentration out to about 100 miles above the surface of the earth.  Humans and altitude typically do not mix well.  Some of our hardier folk live above 10,000 ft, around the great mountain ranges of the world, but they are a distinct minority.  The problem is we tend to get sick at high altitude, especially if we live at low altitude then ascend too fast or stay high too long. 

Charles Houston spent a lifetime researching mountain sickness, which is caused primarily from a lack of oxygen or hypoxia.  Charlie should know all about hypoxia:  he was the leader of two American teams to attempt climbing K2, the second highest mountain in the world at over 28,000 ft.  Both attempts on either side of World War II were unsuccessful but the attempt in 1953 achieved fame for the self-less acts of all climbers involved in the fatal expedition.  The team became known as the brotherhood of the rope and is the subject of numerous mountaineering articles. After this last expedition, Charlie retired from mountaineering and spent a lifetime studying the physiological effects of hypoxia from a number of institutions, including the University of Vermont, where he was a Professor of Medicine.  His article in Scientific American on hypoxia is a great read [1].


One of the physiological effects of hypoxia includes an increased flow of blood to the brain, which creates a local accumulation of fluid and pressure leading to headaches and impeded judgement.  I experienced this on my climb on Kilimanjaro at about 14,000 ft at the famed Lava Tower after a day of about 4,000 ft elevation gain.  This particular day of my trek was carefully arranged by our guides to allow for acclimatization, which helps the body adjust to elevation gain.  The physiological effects include an up-regulation of the hormone erythropoietin to stimulate the production of red blood cells and allow for oxygen to be carried around the body more efficiently.  This important acclimatization step certainly helped with me being able to summit Kilimanjaro without ill effects.  For more on this adventure and some great photographs, please go to the SLAS Electronic Laboratory Nieghborhood.

Peculiar image of a table setting for 1 for lunch that we passed on our way to Kilimanjaro's Lave Tower (background).

1. Houston, C. S. 1992. "Mountain sickness." Scientific American, October, pp. 34-9

SBFC 2012

The 2012 New Orleans Jazz & Heritage Festival was in full swing in the "Big Easy" when the 34th Symposium on Biotechnology for Fuels and Chemicals opened at the Sheraton on Canal Street next to the French Quarter in New Orleans. The meeting, which ran from April 30- May 3, had approximately 700 attendees from around the world who gathered to exchange information on biofuel. There were over 100 oral presentations and 400 posters given over a period of four days. The days were long, with meetings starting at 8:00 AM and poster presentations finishing up at 9:00 pm. Fortunately there was at least one afternoon free to enjoy the sights and sounds of this culturally diverse city. David Glassner from Gevo gave the Keynote address talking about the process of commercialization of biochemicals and how the use of biofuel has to follow the same model as conventional commercialization where a highly efficient cost effective large scale process is defined prior to the pursuit of biological solutions. The implication of which is that many investigators attempt the reverse by trying to fit a developed process into a business case. The meeting had multiple tracks running parallel with 18 different sessions, covering bacterial science and technology, biomass physicochemical analysis, enzyme science and technology, biomass treatment, algae, yeast, biofuel economics and many more. BioTek was one of 20 vendors exhibiting instrumentation and equipment useful for biofuel research and production. In addition, BioTek’s application group presented a poster describing the use of microplate technology for the screening of algal cell cultures.

The French Quarter of New Orleans

By Paul Held, Laboratory Manager

Making Science Interesting and Relevant

As an application scientist for BioTek instruments my typical day involves performing experiments, writing application notes, talking with people, and testing prototype instrumentation; all very interesting things, but not often relevant to my life outside of BioTek. However, when I come across a topic that meshes with my hobbies I get really excited. Recently I have worked on several biofuel applications, which include the digestion of cellulose, xylan, and starch. The optimization of each of these reactions, while interesting, is not something that one would think is relevant to my day to day existence. That is until you consider the purpose of those enzymatic reactions, which is to generate glucose that can then be fermented by yeast into ethanol. This is something that I can relate to, as I also enjoy brewing beer. Brewing, as you may know is the act of extracting and digesting starch from grain to produce sugars (glucose) that are then fermented by yeast to produce the ethanol containing beverage, beer. Eureka… an application note on the production of ethanol in beer! It is not often when my BioTek world and my world outside of BioTek are aligned so well.

Figure 1. Glass fermentation carboys. Small batch brewing methods often employ the use of glass containers for the fermentation process.

Vienna style lager beer was produced by standard production procedures. Briefly, crushed malted barley was wetted and incubated at 77° C for approximately 60 minutes to convert grain starch to glucose by action of the endogenous α-(1→4) and α-(1→6) glycosylase enzymes present in the germinated barley. The sugar rich aqueous extract was isolated by flow through filtration and boiled for 60 minutes. Hop buds were added at intervals during the boil for flavor. After cooling, the unfermented wort was inoculated with Bavarian lager yeast (Wyeast strain 2206). The culture was sealed with an air lock and allowed to ferment at approximately 16° C. Aliquots (15 mL) were removed daily, centrifuged at 800x g and the supernatant stored at -20°C until assayed for ethanol and glucose content.

Figure 2. Vienna style lager fermentation. Glucose is consumed by the yeast Saccharomyces cerevisiea, while ethanol is produced as a byproduct.

During the fermentation process the production of ethanol in fermenting beer was monitored. As demonstrated in Figure 2 the ethanol concentration of fermenting Vienna lager style beer increased steadily from 0 to approximately 5.5% over a period of 350 hours (14 days). This is in good agreement with ethanol calculations based on the change in specific gravity. The initial specific gravity of the unfermented beer was determined to be 1.055 g/mL, while the final gravity was 1.010 g/mL. Using the change in gravity one can estimate the concentration of ethanol produced to be 5.9%. At the same time glucose was being consumed. Glucose is present in high concentration (approx 80 mM) in the beer at the beginning of the fermentation process and is completely consumed in the first 50 hours of fermentation (Figure 2). This rate is much faster than the production of ethanol, suggesting that glucose transporters are quickly sequestering the glucose from the media. Only as the glucose is consumed for energy by the yeast is ethanol produced.

This application is an example of making science interesting and relevant to me, in other words it was fun. What are some examples of that science projects that you would find interesting and relevant? The U.S. Department of Education’s (DOE) National Assessment of Education Progress (NAEP) report indicates that U.S. students are doing poorly in science. I will not bore you with the breakdown of the statistics, as I am sure most will agree with the overall assessment. However, if there were more opportunities where children could be exposed to science that was interesting and relevant (i.e. FUN) to them, then some of the issues with science education might be solved.


By Paul Held, Laboratory Manager

Service Team

Every 4 months (January, May and September) we gather our service teams together to review our operations to date, work on improvement projects together as well as learn about the next wave of products to be released. We try to have our January meeting somewhere besides Vermont because having our U.S. field service team travel up North can be challenging at times in the winter. Also, with our field service team spread around the country it can be as cost effective to have a meeting in a major metropolitan area. And let’s face it, they do a great job so why would we want to reward them with a deep freeze up north?

So this January, actually Jan 31 through February 2, we met our domestic field service team in New Orleans. The days were very busy with discussions about: our successful 2011 results; 2012 targets; and technical training with a deep focus on the new 405 Touch. While down in the Big Easy we were able to meet up with the BioTek regional sales team for a great dinner. It is hard to find a bad meal in New Orleans. One night we also caught a deal and watched a New Orleans Hornets game against the Phoenix Suns.

Right now we have a team of service engineers getting ready to head over to Poland to provide two weeks of technical training to our European teams. The team will return and have just enough time to catch their breath before everyone comes back to Vermont for the second meeting of the year. This meeting will have the most technical content of any meeting to date considering the exciting releases BioTek has planned for 2012.

By Sean Jordan, Service Director

Antibody-Dependent Cellular Cytotoxicity (ADCC) Assays-The Present and The Future

The success of biologic therapeutics has begun to reshape today’s pharmaceutical market. The first and most successful of these antibody therapies, Rituximab (Rituxan®; Roche/Genentech), showed worldwide sales in 2009 of $5.6 billion (GEN News Highlights, 2011). This, among others including Trastuzumab (Herceptin®; F Hoffman-La Roche), have shown great promise for treatment of patients with leukemia, lymphomas, breast, and other cancer types due to their specificity and reduced side effects (Zhou, 2007). One of the mechanisms which play a central role in the response to clinical antibody therapy is antibody-dependent cell-mediated cytotoxicity (ADCC) (Wang, 2008). This involves the response of natural killer (NK) cells to bind to specific antibody-coated target cells, such as CD20 and HER2 expressing cells, to promote the death of the target cell.

With many of the existing patents covering these treatments set to expire in the next few years, the development of biologic therapeutics similar to the original drug (biosimilars) has become increasingly important. This is highlighted by the report that Spectrum Pharmaceuticals and Viropro are set to work together to develop a biosimilar to Rituximab (GEN News Highlights, 2011). As a direct result, assays that can assess the ability of a biosimilar to act in a manner similar to the original biologic have also seen increased interest. The current “gold standard” ADCC assay incorporates 51Cr. The procedure involves labeling and incubating target cells with the radioligand, assessment of the labeling procedure, and finally performance of the actual assay. Not only is this time consuming, but involves the use and eventual costly disposal of radioactive material.

A number of new cell-based technologies have been, and continue to be developed that are easier to use than 51Cr, less time consuming, and do not include the use of radioactivity. These include high-throughput methods that separately assess antibody binding to target as well as CD16 receptors, as well as medium-throughput methods that assess this binding in one assay using genetically engineered cell lines or purified NK cells from blood. Each has its own strengths, and can be the assay of choice depending on the needs of the researcher. In addition, these new assay chemistries are amenable to automated processing and can be detected using microplate readers. This can serve to further simplify the procedure and make a more robust process. This was initially demonstrated through multiple poster and oral presentations at the recent SLAS Conference, and will continue to be illustrated through our presence at a number of important upcoming biological therapeutic focused conferences.

How do you see biological therapeutics continuing to reshape the drug discovery process? What type of bioassays would you like to see automated in your lab?


By Brad Larson, Principal Scientist

Lab Manager: what it takes to be “The Guy”

One of my roles at BioTek is “Lab Manager” of the Applications Laboratory. No I’m not a manager to any personnel, but rather I manage the lab facilities. While an unappreciated endeavor, every lab needs a manager. In the smallest of labs, (i.e. one person), the scientist is also the manager. In very large labs they often will have a person, often referred to as “The Guy”, solely dedicated to the management of lab facilities. Most laboratories have someone in the middle, where one person, either formally or informally, is in charge of the day to day operation of the lab. This is the case here at BioTek, where my main role is that of an Application Scientist, but I am also the Lab Manager. While the job is not without its headaches it is also a lot of fun.

The role Lab Manager is a cross between an accountant and an auto mechanic. Besides maintaining some sort of lab budget and expense ledgers, one also is responsible for instrument repair and maintenance. Besides coordinating equipment usage among different researchers (including myself), the lab manager has to purchase, find space and install new equipment. This lets me get my hands on everything in the lab and for a gadget person that is almost as good as sliced bread. Even the mundane portion of the role of lab manager, such as ordering of supplies has an upside in that you have your finger on what everyone in the lab is doing.

Coordinating schedules is the most daunting of tasks. Routine maintenance of equipment, which often takes things off line, has to be coordinated with everyone’s ever changing need for that equipment. Installation of new equipment may require the interaction of scientists, electricians, plumbers, movers, and facilities personnel. Sometimes it has the feeling of trying to herd cats. But all is forgotten when you fire the new piece of equipment up for the first time. Just like a getting a birthday or a Christmas present.

Even with the best of preventative maintenance, equipment fails from time to time. Failure of instrumentation always seems to take place at the most inconvenient of times, often at night. Either it takes place when that device is most needed by the lab or when you have a number of other equally important tasks to finish. Despite the annoyance caused by equipment down time, getting things back to normal provides a sense of accomplishment (and relief) for getting the job at hand done.

Over the years I have noticed that Lab Manager by committee does not seem to work. Regardless of the size of the lab that I have worked in; I’ve found that it is always best that one person plays this role. In the case of the BioTek Application lab, I’m glad it’s me.



By Paul Held, Laboratory Manager