Improved Method for the Rapid Analysis of Monoclonal Antibodies Using IdeS

ides_abTherapeutic monoclonal antibodies (MAbs) are inherently heterogeneous due to a wide range of both enzymatic and chemical modifications, such as oxidation, deamidation and glycosylation which may occur during expression, purification or storage. For identification and functional evaluation of these modifications, stability studies
are typically performed by employing stress conditions such as exposure to chemical oxidizers, elevated pH and temperature.

To characterize MAbs, a variety of analytical techniques are chosen, such as size exclusion chromatography and ion exchange chromatography. However, due to the large size of the intact MAbs, these methods lack structural resolution. Often, the chromatographic peaks resolved by SEC and IEC methods are collected and further analyzed by peptide mapping to obtain more detailed information. Peptide mapping, in which antibodies are cleaved into small peptides through protease digestion followed by LC–MS/MS analysis, is generally the method of choice for detection and quantitation of site-specific modifications. However sample preparation and lengthy chromatographic separation make peptide mapping impractical for the analysis of large numbers of samples. In contrast to peptide mapping analysis, the middle-down approach offers the advantage of high-throughput and specificity for antibody characterization.

Limited proteolysis of IgG molecules by the IdeS enzyme has been introduced for antibody characterization due to its high cleavage specificity and simple digestion procedure. Continue reading

You Become Your Habits

29980706-AugustBlog-BrainPuzzleHumans are, by nature, creatures of habit. The Experts Blog from the Cleveland Clinic tells us up to 95% of our thoughts are repeated every day–habitual thoughts, and they can take root in our lives without our conscious realization. Habits help us get through the day. Imagine having to wake up each morning and decide what you should do next. Knowing that you need to shower, eat breakfast, brush your teeth, etc. saves us some major brain power. Problems can occur when our habits unknowingly become incongruent with our values and goals. Many of us have habits that no longer serve us, such as smoking, the way we respond to our spouse or the thoughts we have about ourselves. These habits can keep us stuck and they don’t leave much room for creative thought and solving problems.

The obvious question is “how can we break free from obstructive habits?” Committing to just 10-15 minutes of meditation per day will create changes in the brain that allow you to increase the time between stimulus and response, which will give you greater freedom in how you respond and react. The cultivation of conscious awareness through meditation, therefore, can help us break habits that are unhealthy or that are no longer serving us. Just by recognizing our habits and placing our attention on them they will begin to soften. With commitment we can begin to see these habits change.

If you want to try an experiment, pay attention to the choices you make and your reactions over the next few days. Make a mental note of these thoughts and reactions and decide if they are in line with your values. You may also begin to notice what triggers precede these habitual responses.  If they are thoughts or actions you’d like to change, make an intention to respond differently next time. No one can break these deeply rooted patterns all at once, but by noticing and deciding what you’d like to do differently, you will be taking a huge step forward in the process.

How’s That for Wellness?

Whether you’re a seasoned runner or just trying to find a fun activity to do with a coworker, a 5K run/walk can be a fun event for all.  Promega recently hosted its 7th annual 5K run/walk and had more than 155 participants at the Madison, WI,  location.  The event was a fun activity for everyone to get out, get active and be social. The event also helped raise over 160 pounds of food and over $115 in cash that was donated to the Second Harvest Food Bank.

As a nation, we have become more focused on our health and wellness than ever before. Nearly everyone is trying to drink more water, be more mindful of what they eat, and it’s hard to look around the room without finding someone wearing a fitness tracker to keep track of their daily steps.  Health and wellness has also become increasingly popular in the workplace. According to Health Fitness Revolution, the top benefits of a Workplace Wellness Program are as follow:

  • Fun
  • Improved Productivity
  • Happiness
  • Sense of Community
  • Lower Healthcare Costs
  • Sense of Accomplishment
  • Improved Physical Fitness
  • Weight Loss
  • Less Stress
  • Healthier Habits

At Promega our focus on wellness as an organization is no different. We recognize that being active and taking care of your health, and your family’s health, has an impact beyond how you feel.  Because of that, we have a wellness or fitness center in each of our facilities to make staying active and fit more convenient. We also believe that wellbeing extends beyond physical health and have a campus that features native gardens with walking paths and dedicated meditative spaces to encourage total wellness. Promega strives to be a leader in health & wellness initiatives that enable our employees to become the best version of themselves. We don’t do this for a potential monetary return on the investment, but because we truly believe that by becoming the best version of yourself, our employees also become the most fulfilled.


ViaFect™ Reagent for Transfection of iPSC-derived Cell Lines

Madison, WI is home for Promega, and while it is not a huge city, Madison is home to many biotech companies, fed mostly by the local, world-class University of Wisconsin-Madison. Many scientists and scientist families work, live and play near one another here. It is not uncommon for two scientists from different companies to talk to one another and discover that their respective companies have products or processes that could benefit both companies.

Case in point: Scientists at Promega have a good working relationship with Cellular Dynamics International (CDI), a biotech firm that specializes in differentiated iPSC-derived cells. We want to demonstrate that our assays work in iPSC cells and CDI wants to demonstrate the range of assays that can be performed with their iPSC-derived cells.

Differentiated iPSC cells are as close to primary cells as you can get, and primary cells are notoriously difficult to transfect due to their slow rate of growth and increased propensity for death. CDI reported great success with ViaFect™ Reagent and generously shared their data with us (see image). Continue reading

What’s Art Got to Do with It?

While some may see the Art Showcase that Promega has sponsored for the past 20 years as tangential to the mission of the biotechnology company, these quarterly exhibits of local and global artists contribute to Promega’s commitment to creativity and innovation in the arts, culture and sciences. The exhibits also foster connections between members of the community that probably would not otherwise exist.

Promega President and CEO Bill Linton with Daniel Swadener, curator of the Promega Art Showcase, at the 2016 Fall Art Showcase Opening and Symposium.

Promega President and CEO Bill Linton with Daniel Swadener, curator of the Promega Art Showcase, at the 2016 Fall Art Showcase Opening and Symposium.

It is obvious how the show serves to advance the arts and culture, but its relationship to science is less clear. Based on my experience attending the symposium and viewing the artwork, the science at Promega benefits from this endeavor as well.

Let me begin by describing the work included in this fall’s Art Showcase, “Wis-Con-Sin.” This exhibit features three centuries of Wisconsin photographers that each created life-long photographic projects based in Wisconsin:

  • Charles Van Schaick (1852-1946) was a studio photographer in Black River Falls, WI who left behind nearly 6,000 glass plate negatives of mostly studio portraits (which have been featured in two books, Wisconsin Death Trip and People of the Big Voice), as well as street scenes, major events in the region, outdoor family and group photos, buildings, picnics, people and livestock.
  • Eugene Von Bruenchenhein (1910 – 1983) was a self-taught artist who created several thousand works including apocalyptic oil paintings, ceramic crowns and vessels, and photographs that he and his wife Marie collaborated on, staging her in provocative poses and costumes.
  • J. Shimon & J. Lindemann collaborated as artists since 1983, focusing on rural Wisconsin towns where they both grew up and using antiquarian cameras and printing techniques to record post-industrial settings, rural landscapes, small towns, and shifting modes of life.

Continue reading

Back to Basics: Organizing Your Writing like It’s a Hamburger

The "hamburger" scheme for organizing a paragraph.

The “hamburger” scheme for organizing a paragraph.

Last night I was helping my daughter, who is in fourth grade, with her homework. We had completed a math worksheet, a geography worksheet and had moved onto writing. For her paragraph assignment, she was supposed to write about a special place. So I began drawing the concept map that we typically use to help her organize her thoughts. She stopped me before I could get started.

“No Mom, wait,” she grabbed the pencil and paper from my hands, “I have a better idea.”

She drew five shapes on the paper.

“We should write the paragraph like it’s a hamburger. The first sentence is the topic—it’s the top of the burger, tells you what is inside—it makes you hungry to read more. Next comes the juicy, meaty part. Three details—three sentences. Then the bottom bun, the summary that supports the whole paragraph. It’s the hardest to write.” She proudly sat down with her drawing and pencil.

“I LOVE that,” I exclaimed. “That’s a great way to organize a paragraph.”

“Yeah,” my husband looked up from his Suduko that he had been working on, “and the cheese goes right here.” He pointed to one of the three boxes my daughter had drawn underneath the bun.

“And the lettuce over here,” my daughter giggled.

“Well, I like mine with lettuce and tomato,” I chanted with no apologies to Jimmy Buffett, “Heinz 57 and French-fried potato..,”

“A big kosher pickle,” my daughter joined in, and the evening’s homework activities degenerated from there. (Sometimes it’s the parents who are easily distracted.)

My daughter’s hamburger graphic was new to me, but the concept wasn’t. It is a solid method for organizing a piece of writing, and it can be applied all kinds of writing—from a paragraph, to an essay, to a speech and even to a scientific article. Continue reading

We Need More Conferences Like ISHI: Memoirs of a First Timer

Keynote speaker David O’Shea kicked off the ISHI 27 conference with his investigation on how DNA profiling is helping to reunite families in Argentina after the military coup in the late 70’s.

We shared in laughter and tears. We tempered our scientific pursuit of the truth with the story of an unimaginably strong survivor of rape. We witnessed the struggles of a man trying to find his identity and the joy of being reunited with real family members after 30 years of lies. I find it hard to succinctly describe to others what my first ISHI conference was like. There is perhaps nothing more personal than our own genetic identities. This conference didn’t shy away from the raw emotions that encompass the human experience. We define ourselves as employees of this company or researchers at that institution, competing for attention and funding, yet this conference reveals how limiting these preconceptions may be.

The desire to make the world a better place unites us. I spoke with analysts for hours about the challenges of overcoming the sexual assault kit backlog, I made a fool of myself dancing to musical bingo with new friends from the Philippines and Brazil, and I was inspired by the casual musings of a video journalist. We are sure to see countless more ethical debates on how we should be using DNA (or proteins!) for human identification. The field of science relies on the open sharing and exploration of new ideas, and as admittedly biased as I am to the conveniences of the digital age, there has never been a better time to come together in person.

Don’t just take my word for it, though.

There were some phenomenal talks each day, and I did my best to capture the essential takeaways from Continue reading

Of Elephant Research and Wildlife Crime – Molecular Tools that Matter

Here at Promega we receive some interesting requests…

Take the case of Virginia Riddle Pearson, elephant scientist. Three years ago we received an email from Pearson requesting a donation of GoTaq G2 Taq polymerase to take with her to Africa for her field work on elephant herpesvirus. Working out of her portable field lab (a tent) in South Africa and Botswana, she needed a polymerase she could count on to perform reliably after being transported for several days (on her lap) at room temperature. Through the joint effort of her regional sales representative in New Jersey/Pennsylvania (Pearson’s lab was based out of Princeton University at the time) and our Genomics product marketing team, she received the G2 Taq she needed to take to Africa. There she was able to conduct her experiments, leading to productive results and the opportunity to continue pursuing her work. Continue reading

Next-Generation Genomics Education: Educating and Preparing the Next Generation

Students pursuing their interests wit h hands-on activities at BTC Institute.

Students pursuing their interests wit h hands-on activities at BTC Institute.

The BTC Institute has many partners in creating educational opportunities in the molecular biosciences. In recent years, we have worked with the Dane County School to Work Consortium (DCSWC) to create a unique, one-semester class aimed at giving high school Juniors and Seniors interested in scientific research and health careers a chance to explore how concepts they have been learning about in their biology and biotechnology classrooms are used in the laboratory.

To take it one step further, these laboratory experiences are tied to the Gates Foundation Grand Challenges in Global Health. The Challenges provide a framework to help students understand worldwide concerns, including ways in which biotechnology can be applied to generate solutions to these problems. Students are encouraged to place scientific challenges within social and socioeconomic contexts which, in turn, make some solutions more appealing than others. This holistic approach provides the “real world” milieu that is so sought after in academic endeavors. Continue reading

Friday Cartoon Fun: Take My Samples, Please!

Instruments can make our lives easier in the lab. Place your samples inside an instrument and let it do all the work—isolating nucleic acids or reading and analyzing a multiwell plate—while you walk away to read a new research paper or prepare for the next step in your experiment. However, with the array of machines now available to scientists worldwide, some confusion may result in the laboratory. Has this ever happened to you?


Copyright Ed Himelblau