What Does This Do? HPLC

Recently, I have been using a machine called a HPLC quite a lot in my research. This has lead to quite a lot of questions like, “What is an HPLC? What does it actually do?” mostly asked by my grandma.

So, HPLC stands for High-Performance Liquid Chromatography, which is a mouthful. One will also hear it referred to by an older term, High Pressure Liquid Chromatography. You can see why most scientists are lazy and just refer to it as HPLC.

Erin Gallagher at the HPLC.

Erin Gallagher at the HPLC.

What a HPLC actually does is force a liquid mixture, which you want separated, through a tube of packed beads (called a column) at high pressure. In this liquid mixture there is some component that you want to separate from the rest of the mixture, whether it is a protein you need purified after synthesis or a drug from a urine sample. This is how doctors monitor that you are getting the right dosage of a drug and one of the ways that cocaine and other illicit drugs are tested for1.

As the sample passes through the column certain components in the sample will be attracted to the packed beads. Those components will take longer to get through the column because they keep getting stuck to the beads as they go through the column. This means that some components in the mixture will fly through the column, while others will take much longer to get through the column because those components keep sticking to the beads and then unsticking. This process is how the HPLC separates the mixture into the different parts. The sample can be separated using size, polarity, or several other chemical properties.

A detector is attached at the end of the column to identify what is coming off the column when. The detector can use many different types of detection, from ultra violet/visible light to mass spectroscopy, to figure out what component of the mixture is coming off of the column at what time.

Overall, the HPLC helps scientist separate and identify, and sometimes even quantify, parts of a liquid mixture.

1) Heit et al. Urine drug testing in pain medicine. Journal of Pain and Symptom Management March 2004. Pages 260–267


For a more thorough walk through and some awesome diagrams of HPLC see:

Harris, Daniel C.. Exploring Chemical Analysis. 4th ed. New York: W. H. Freeman and Company, 2009. Print.


Erin Gallagher is a second year PhD student in Peter Searson’s Materials Science and Engineering lab.

ChemBE seminar focuses on cancer research innovation

The Department of Chemical and Biomolecular Engineering’s  scheduled Thursday, October 10 seminar will continue as planned at 3:30 PM in Maryland Hall 110. Jerry Lee, the Health Sciences Director at the National Cancer Institute (National Institutes of Health) will present his lecture “Advancing Convergence and Innovation in Cancer Research: National Cancer Institute Center for Strategic Scientific Initiatives (CSSI).”  A small reception will follow in Maryland Hall 109.

Jerry Lee

Jerry S.H. Lee, Ph.D


The National Cancer Institute (NCI) Center for Strategic Scientific Initiatives (CSSI) is a component of the NCI’s Office of the Director focused on emerging advanced technologies that have the potential of uniquely impacting the full spectrum of cancer basic and clinical research. The Center is tasked with planning, developing, executing, and implementing rapid strategic scientific and technology initiatives that keep the Institute ahead of the scientific curve with respect to potential new exciting areas and discoveries. This may involve direct development and application of advanced technologies, synergy of large scale and individual initiated research, and/or using available federal mechanisms to forge novel partnerships that emphasize innovation, trans-disciplinary teams and convergence of scientific disciplines. With an emphasis on complementing the scientific efforts of other NCI divisions, CSSI’s efforts seek to enable the translation of discoveries into new interventions, both domestically and in the international arena, to detect, prevent and treat cancer more effectively. This presentation will highlight various programs and their associated accomplishments within CSSI’s broad scientific portfolio of programs (Clinical Proteomic Tumor Analysis Consortium, Alliance for Nanotechnology in Cancer, Physical Sciences-Oncology Centers, Innovative Molecular Analysis Technologies, and Provocative Questions) and describe future directions and opportunities.

Bee blight and a honeycomb of funding possibilities

by Holly Occhipinti http://www.flickr.com/photos/pinti1/5519458297/

by Holly Occhipinti http://www.flickr.com/photos/pinti1/5519458297/

Time Magazine had an issue recently where the front cover asked, “What would the world be like without bees”? (1) This might seem like a rather small loss to the global ecology; how could bees play that large of a role in our daily lives? Actually, bees are rather important for us as human beings and for our food sources. The article estimated that the loss of bee pollination of plants would cost several billion dollars annually. This is perhaps the reason why a funding source we as scientists might not be accustomed to thinking about, the U.S. Department of Agriculture through the Agricultural Research Services (ARS), is looking to fund research to protect bees. (2) I believe that this could provide a unique source of funding for labs interested in studying and preventing the spread of Bee Blight.

What is Bee Blight then and why is it threatening bees?  As the source from above writes, Bee Blight is a poorly understood phenomenon, which is causing a change in worker bee behavior to have them leave their hives.  This decreases the overall population of bees and the effectiveness of the bee hive at producing honey and pollinating.  It is speculated that a combination of pesticides and fungicides that are currently in use are somehow poisoning the honey bees and causing them to exhibit this strange behavior.  Due to the fact that this concepts is very poorly understood, there are a multitude of efforts being made to find solutions to the problem of Bee Blight.

We as scientists at Johns Hopkins University have an opportunity to help research this problem and, in terms of funding, gain access to the resources that the ARS and other groups is providing.  Recently, a Harvard Lab was provided almost ten million dollars from the NSF to study how to make a hive of robotic bees to replace the ones found in nature. (3)  As someone whose research project is based on studying Melittin, the key component of Bee Honey Venom, and who is also an amateur entomologist, I believe that the solution might not need to be that exotic.  How many of us work on nanoparticles that are supposed to have very specific properties in very specific environments?  How many of us work on creating coatings for devices that kill off bacteria and other harmful diseases?  How many of us work in the public health department and know the impact that proper food management has on people’s health?  I believe that there are many other options, from better crops to better pesticides, that we at Hopkins have the ability to investigate and that this provides a new source of funding for our efforts.

  1. Time Magazine, August 2013
  2. Kaplan K. (2013, March 7). Honey Bees and Colony Collapse Disorder.  ARS website. Retrieved 9/9/2013 from <http://www.ars.usda.gov/News/docs.htm?docid=15572>
  3. Robobees Lab. (2013). Home Page. Robobees Lab Website. Retrieved 9/9/2013 from <http://robobees.seas.harvard.edu/>

By Gregory Wiedman, a graduate student from the Materials Science Department who is altering natural peptides from Bee Honey venom to improve drug delivery.

Info sessions on international research internships

IMEC clean room

What’s better than working on a cool research project in your lab? Why it’s working on a cool research project in a fascinating European country, of course!

Johns Hopkins Institute for NanoBioTechnology offers undergraduate and graduate research internships at IMEC’s world-class nanofabrication laboratory in Belgium. Internships last approximately 10 weeks and include housing and a stipend. Find out how to apply and what kinds of projects are being sought at one of our upcoming informational sessions. Two sessions will be held October 8, one at 1 p.m. with light refreshments and a second at 5 p.m. with pizza, both in Croft Hall, Room 170.

RSVP is required to Tom Fekete at tfekete1@jhu.edu

Take a nanobreak at INBTea Time every Wednesday

Johns Hopkins Institute for NanoBioTechnology is pleased to announce the first INBTea Time every Wednesday in the corridor outside of our headquarters at 100 Croft Hall from 2:30 to 3 p.m. Grab a drink and a light snack. The first INBTea Time will be held on Wednesday, September 11. Enjoy a quick break and great conversation!

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Blog posts from Belgium


Catholic University of Leuven (Photo by Colin Paul)

During the summer, a select group of researchers from Johns Hopkins travel to Leuven, Belgium to work at the micro- and nano- electronics fabrication laboratories of IMEC. Usually three to five students are able to go over for up to three months to work on a research project that is collaboratively arranged by both IMEC researchers and Johns Hopkins University faculty.

The students usually are advancing some aspect of a project they have started here at JHU. In a few instances, researchers from IMEC come to JHU to work. Faculty at both locations work together to develop projects that are mutually beneficial to all parties.

Johns Hopkins Institute for NanoBioTechnology provides financial support for our research through the National Science Foundation’s International Research Experience for Students (IRES) program. The arrangement with IMEC has been in place since 2009.

IMEC, which used to be referred to as the Interuniversity Microelectronics Centre, is an unusual research entity that grew out of an agreement between the Flemish government and the academic community at Catholic University of Leuven They now have more than 2,000 researchers from all around the globe working at their high tech facility.

To keep in touch with our researchers while they are away and to find out about their outside of laboratory adventures, INBT established the IMEC blog.  Click here to check out what our students have done, this summer and over the last several years.


Are cellular technologies scalable?

Are cellular technologies scalable? According to Phillip Vanek, Head of Innovation at Lonza Bioscience, the answer to this question is “yes”, but only if scaling is considered very early in the technology’s development. Vanek addressed the topic of scalability at his talk at the INBT symposium.

scalabilityScaling-up of bench-top science into industrial processes is difficult for a number of reasons. Commercial-scale production of cell-based products introduces regulatory challenges and production volumes never encountered on the bench scale. Even the basic laboratory chore of cell passage can become a large hurdle when attempting to grow large number of cells in the multi-layered cell factory system.

With such challenges in mind, Vanek lays out a number of ways to improve the success rate of scaling up processes. He stressed that a process should ideally be closed for maximum success. A closed process prevents product contamination and minimizes user error. Also, maximizing automation helps minimize operator error in processing.

Most importantly, the treatable patient pool sizes and dosage requirements need to be well-known for a process to be commercially successful. Vanek concluded that cellular technologies are scalable, but only if researchers start with end goals in mind early and are well-aware of potential pitfalls.


Editor’s Note: This a summary of one of the talks from the 2013 Nano-bio Symposium hosted by Johns Hopkins Institute for NanoBioTechnology held May 17. This summary was written by Christian Pick, a doctoral candidate in the chemical and biomolecular engineering laboratory of Joelle Frechette. Look for other symposium summaries on the INBT blog.


Johns Hopkins opens mind-blowingly beautiful teaching labs

When I was an undergraduate at [insert state university name here] most of the biology and chemistry teaching labs we undergrads worked in were purely utilitarian spaces for bench work and all had seen better days. In fact, they looked more like my high school science labs.  My high school labs might have been nicer.

New Undergraduate Teaching Labs. (Photo by Mary Spiro)

New Undergraduate Teaching Labs. (Photo by Mary Spiro)

But there is no reason why even lower level science should not be taught in an aesthetically pleasing space. For some undergraduates, lower level science courses are a requirement that must be knocked off the graduation to-do list. Why not make the experience more exciting?

Johns Hopkins University has constructed 105,000 square feet of new undergraduate laboratory teaching space on the far side of Mudd and Levi Halls on the Homewood campus. The space beautifully integrates into the natural surroundings offered by the Buffano Sculpture Gardens. The open feel is extremely important, I think, because it allows so much natural like to come in  that it could make that time in lab section, which can seem never-ending, into something much more tolerable–even enjoyable.

Undergrads studying chemistry, biology, biophysics, neuroscience and psychological and brain sciences will be using the 20 labs that were created with the new space. The philosophy behind the design of the building is to encourage cross disciplinary collaboration, which as many know is something that Johns Hopkins Institute for NanoBioTechnology strives for in all of its endeavors. It is nice to see that same kind of sentiment carried out in a physical way in this new campus structure.

For now, the addition is just being called the UTL (undergraduate teaching laboratories) building. The core of the structure includes space for nuclear magnetic resonance imaging and tissue and cell culture. Lab space connects to the adjacent academic buildings via mingling areas, ample seating, faculty offices, a large biology research lab, a computer lab, seminar rooms, a coffee bar and more. 

I recommend walking over to the UTL yourself and having a look around if you are on the Homewood campus. If not you can watch this video. It will make you want to register for a chemistry class like a freshman. No, joke. This building is a work of art and an amazing addition to the already beautiful Homewood campus.

Read the story about the new laboratory building here.

Commercialization of nanotech no easy task

Editor’s Note: The following is a summary of one of the talks from the 2013 Nano-bio Symposium hosted by Johns Hopkins Institute for NanoBioTechnology held May 17. This summary was written by Christian Pick, a doctoral candidate in the chemical and biomolecular engineering laboratory of Joelle Frechette. Look for other symposium summaries on the INBT blog.

Govt-RegulationsOne of the greatest promises of merging nanotechnology with medicine is in the creation of highly selective vehicles for drug delivery based on nanoparticles. However, translating nanoparticles into commercialized medical products comes with many challenges. Anthony Tuesca, a scientist in the Innovative Drug Delivery Group at MedImmune, outlined a number of these challenges and ways to address them. 

Research in nanotechnology begins at the lab bench, often without much thought given to future commercialization. But commercialization is a huge undertaking. In between the lab-bench and final product there is a whole litany of challenges that must be tackled. One such challenge lies in scaling up processes for production. As Tuesca stated, for scale-up to be viable, laboratory processes must be compatible with current manufacturing capabilities.

Another challenge is navigating an often confusing regulatory landscape. Although nanoparticle based therapies don’t necessarily invoke harsher requirements than conventional medical treatments (in that both require clinical trials that demonstrate safety and efficacy) more complex technology requires more proof of effectiveness. Interestingly, Tuesca mentions that the U.S. Food and Drug Administration currently lacks an official definition of nanotechnology.

Ultimately, Tuesca’s presentation urges researchers to take a proactive role in translating laboratory discoveries into viable medical technology. Such a role requires researchers to consider future commercialization early in their research and act accordingly.

Did you know that the Johns Hopkins Center of Cancer Nanotechnology Excellence has a working group focused on commercialization? Read more about it here.


Unlocking the mysteries of the blood-brain barrier

It might astonish you to know that, although we use our brains all the time, science knows very little about how they actually work. That is why recently, President Barack Obama announced a $100 million initiative to map the human brain.

“We can identify galaxies light-years away; we can study particles smaller than an atom; but we still haven’t unlocked the mysteries of the three pounds of matter that sits between our ears,” Obama said in a press conference on the announcement April 2.

The blood-brain barrier involves functional interactions between endothelial cells that form brain capillaries, astrocytes, and pericytes in a complex microenvironment. (Illustration by Martin Rietveld)

The blood-brain barrier involves functional interactions between endothelial cells
that form brain capillaries, astrocytes, and pericytes in a complex microenvironment. (Illustration by Martin Rietveld)

Obama’s Brain Research Through Advancing Innovative Neurotechnologies (BRAIN) project will seek to discover what occurs between the 100 billion cells firing inside the brain with the goalof helping to prevent and even cure neurological diseases, such as Alzheimer’s or Parkinson’s, that affect as many as 100 million Americans.

Johns Hopkins University is at the forefront of brain science research. The Brain Science Institute (BSi) at the Johns Hopkins School of Medicine was launched to develop new multidisciplinary research teams; create cutting edge-research cores for use by all brain researchers at Hopkins; and foster translation of discoveries to treatments of brain diseases, in part, by improving our ability to partner with industry and biotechnology.

In 2012, Peter Searson, professor of materials science and engineering and director of Johns Hopkins Institute for NanoBioTechnology (INBT), joined forces with Jeffrey Rothstein MD, PhD, director of the BSi, to create the Blood-Brain Barrier Working Group. This group brings together researchers with diverse interests and expertise to address key problems associated with drug delivery, to discover the role of the blood-brain barrier (BBB) in disease, and to elucidate the structure and function of the BBB.

“The blood-brain barrier is a dynamic interface that separates the brain from the circulatory system and protects the central nervous system from potentially harmful chemicals while, at the same time, regulating transport of essential molecules and maintaining a stable environment,” Searson said. “It is formed from highly specialized endothelial cells that line the brain capillaries, which transduce signals in two directions: from the vascular system and from the brain. The structure and function of the BBB is dependent upon the complex interplay between different cell types, specifically the endothelial cells, astrocytes and pericytes, within the extracellular matrix of the brain and with the blood flow in the capillaries.”

Although the BBB serves the important purpose of tightly regulating the environment of the brain and preventing sudden changes, which the brain cannot tolerate, Searson said, “this interface also blocks the passage of drug molecules to treat disease, neurodegenerative disorders, inflammation or stroke. Unfortunately, animal models are insufficient for use in under-standing how the human blood-brain barrier functions or responds to drugs. In addition, little is known about how disease, inflammation or stroke disrupts or damages the blood-brain barrier.”

With this in mind, the BBB working group has two primary goals, Searson explained. “Our long-term goal is to build an artificial microvessel that will be the first platform that recapitulates a brain capillary in its local microenvironment. This will enable fundamental studies as well as drug discovery and the development of methods to cross the blood-brain barrier,” Searson said.

The second goal is to understand how the blood-brain barrier can be damaged or disrupted so that strategies can be developed to repair it. Injury and disease can disrupt the normal structure and function of the blood brain barrier.

Currently the BBB Working Group has 40 researchers from disciplines as diverse as anesthesiology, materials science and engineering, pharmacology and oncology. Three postdoctoral fellows and 12 pre-doctoral students are also involved. The group meets monthly and hosts expert speakers on various topics. The working group website also lists current funding opportunities to which members can apply and conferences and workshops of interest.

Membership in the working group is open to any student, faculty member or staff at Johns Hopkins University in any discipline.

Visit the Blood-Brain Barrier Working Group website here.

This article was written by Mary Spiro and appeared in the 2013 issue of Nano-Bio Magazine.