Wyatt Technology

Developing a Library of Proteins for Drug Research

The abundance of protein drugs on the market shows there is an increasing demand for knowledge within the formulation of biologics, as Professor Pernille Harris, Principal Investigator and Coordinator of the Protein-excipient Interactions and Protein-Protein Interactions in formulation (PIPPI) project explains

What are the key aims of the PIPPI project and what does it hope to achieve?
One intrinsic area of focus is training PhD students in the biophysical and structural characterisation of proteins in a pharmaceutical setting. They will create a strong intersectoral network between academic and industrial sectors. They will be key opinion leaders of the future. Scientifically, the aims are to create a representative library of structurally diverse proteins, and through a systematic study of their microscopic and macroscopic behaviour in different environments, we will obtain a comprehensive dataset. We will create a database enabling how to best predict protein stability on a minimum number of experiments and guide how to select the best experiments. We will bring biophysical understanding and characterisation tools for formulation of biologics to the next level.

PIPPI article

Who will benefit from the research
There will be widespread benefits across academia, industry and the wider public. Global academic research is likely to take a big step forward due to the unprecedented amount of coherent biophysical data that will be made available by the consortium. For industry, it will deliver the ability to recognise the potential values of biophysical techniques normally used only in academia. They will also gain an understanding of which of the currently used high throughput techniques add more value to the drug development process. For society, it will improve healthcare due to more robust formulation development processes, and at the same time, improve the costeffectiveness of biologics. It will also allow the European Union (EU) to build on its strong pharmaceutical industry and take the lead in future drug development. Having the edge in knowledge on protein formulation will lower the risk of failure and increase the likelihood of better and more convenient dosage forms (for example, suitable for home treatment instead of hospitalisation). Overall, industry and academia in the EU will benefit from a large pool of highly skilled researchers and research groups.

Not forgetting his LSU training, Jon faithfully introduced Nicole to Plate Reader operation and data analysis, including a complete tutorial on the interpretation of autocorrelation functions (ACF). Apparently the ACF analysis made a great impression! Nicole was hooked on the DynaPro and has since published results obtained with it in Biomacromolecules, Advanced Biosystems, Nanoscale and Molecular Pharmaceutics.

Why do so few universities in Europe have formulation of biologics as a subject?
Except for a few examples, the use of biologics is a relatively new field within the medicinal industry. Traditionally, the big pharma industry is focused on small molecules, and teaching in academia, as well as public funding, are often driven by industrial opportunities. In society, it is recognised that protein drugs are more expensive and difficult to develop and less convenient for the patient, whereas it is not acknowledged that at the same time, biologics are often safer, with fewer sideeffects and are much more precise in their mechanism of action and efficacious in treatment of diseases compared to small molecule-based drugs.

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Featured Customer: Jon Pokorski

Nanoparticle sizing, protein buffer optimization for crystallization, biotherapeutic stability evaluation, critical aggregation or micelle concentration. These are some of the more common applications and uses of the DynaPro® Plate Reader, including the just-launched Plate Reader III. But, some of our Plate Reader customers are far more creative than the average scientist.

Prof. Jon Pokorski, now an Assistant Professor in the Department of Macromolecular Science and Engineering at Case Western Reserve University, was an avid user of the DynaPro Plate Reader during his postdoctoral stint at Scripps Research Institute in La Jolla, CA. His primary use was the characterization of viral nanoparticles used as delivery vehicles for cancer therapies, and the instrument factored into his 2011 publication “Cell Targeting with Hybrid Q Virus-Like Particles Displaying Epidermal Growth Factor” Pokorski et al. (2011), ChemBioChem 12(16), 2441-2447.

Jon Pokorski

Prof. Nicole Steinmetz, now the George J. Picha Designated Professor in Biomaterials at Case Western Reserve University School of Medicine and Director of the Center for Bio-Nanotechnology, was also a postdoctoral researcher at Scripps, working on the engineering of viral nanoparticles and their potential uses for drug delivery and medical diagnostics. Like Jon, Nicole conducted her research under the tutelage of Dr. MG Finn. Finn asked Jon to train Nicole on the use of the DynaPro Plate Reader in order to quickly size her nanoparticles.

Not forgetting his LSU training, Jon faithfully introduced Nicole to Plate Reader operation and data analysis, including a complete tutorial on the interpretation of autocorrelation functions (ACF). Apparently the ACF analysis made a great impression! Nicole was hooked on the DynaPro and has since published results obtained with it in Biomacromolecules, Advanced Biosystems, Nanoscale and Molecular Pharmaceutics.

One thing led to another, and Jon and Nicole found that they had more in common than just a love of dynamic light scattering and ACF interpretation. At first it was a common fondness for outdoor sports such as snowboarding, surfing, hiking and camping; then a love of each other. Last month they were married, culminating the match made in DynaPro heaven. Wedding photos available at http://www.pokometz.com/. The matchmaker Plate Reader was not invited, but took no offense.

Having solved the famous ‘two-body problem,” moving on to co-located positions with Case Western Reserve and a Möbius for determination of size and zeta potential of nanoparticles, Professors Steinmetz and Pokorski continue the light scattering romance. Asked “Did you, or do you ever discuss light scattering… outside of the lab?” Jon’s reply was: “That may be a bit too nerdy for us, but I can’t say that it has never come up.” We look forward to the continuation of successful partnerships with both of them and are on the lookout for more matchmaking opportunities.

Plate Reader, Plate Reader, make me a match!

Jon shared his plate reader love story in the Q&A that follows

What are your and your fiancée’s backgrounds (School, studies, and hobbies)? What led you both to end up where you are today?

Jon Pokorski: I’m a Cali guy and have a BS in biochemistry from UCLA, PhD from Northwestern in organic chemistry, then we both met doing our post-docs at The Scripps Research Institute in San Diego. My advisor, MG Finn, asked me to train Nicole on the plate reader and that was that! We both share a love of outdoor sports: snowboarding, surfing, hiking/camping, etc.
Nicole Steinmetz: From Essen Germany. The following blurb is from her website. Dr. Steinmetz trained at The Scripps Research Institute, La Jolla, CA where she was a NIH K99/R00 awardee and AHA post-doctoral fellow (2007-2010); she obtained her PhD in Bionanotechnology from the University of East Anglia where she prepared her dissertation as a Marie Curie Early Stage Training Fellow at the John Innes Centre, Norwich, UK (2004-2007). Her early training was at the RWTH-Aachen University in Germany, where she obtained her Diploma (Masters) in Molecular Biotechnology (2001-2004) after completing her pre-Diploma from the Ruhr University Bochum, Germany (1998-2001).

How did you first come into contact with the Plate Reader and if you are currently using the Plate Reader what are you using it for?

Interestingly, we had an old Wyatt MALS that I was trying to rehab in the lab and took the LSU training at Wyatt in 2007. Turns out that the old instrument was kaput, and we replaced it with the plate reader. At the time, Nicole and I both were using the instrument for sizing viral nanoparticles that we were using as delivery vehicles for cancer therapies. We still use Wyatt instruments for that purpose today!

How did you and your fiancée meet? Was it really over the plate reader? If so, what are the details!

I believe, that this was in fact the first time we ever had a conversation. I trained her how to use the instrument, complete with tutorial over autocorrelation functions. Me, being the putz I am, took an additional 4 years to realize she was interested in things other than particle sizing!

Do you both use the plate reader now? If so, what is your fiancée researching?

We both use Wyatt instruments but no longer the plate reader (sadly). We use the Mobius for the same purposes, sizing and zeta potential of viral particles. These are manly used for potential treatments in cancer.

Did you, or do you ever discuss light scattering with your fiancée outside of the lab?

That may be a bit too nerdy for us, but I can’t say that it has never come up.

Did you win over your fiancée because of your impressive plate loading abilities, or was it something else about you that caught her eye?

I like to think that it is my ability to interpret autocorrelation functions!

Feel free to share any additional thoughts about anything that wasn’t covered but you feel is necessary to the story!

Perhaps you may want to see the wedding website and our personal sites for the lab.

Featured Customer: John Matson

Prof. Matson’s research focuses on the synthesis of unique polymeric and supramolecular materials with applications in signaling gas delivery, tissue engineering, and sustainability. Current areas of interest include the development of materials for delivery of hydrogen sulfide (H2S), a vital biological signaling gas, the synthesis of cone-shaped bottlebrush polymers, and the preparation of new polymers and polymer blends derived from renewable feedstocks.

Prof. Matson has used Wyatt instruments since his graduate school days in the lab of Bob Grubbs at Caltech. He attended LSU in July 2005. His group currently uses a HELEOS II and T-rEX for absolute molecular weight determination for a variety of polymers, including complex cone-shaped structures.

John Matson

“It’s been over a decade since I attended LSU, but I still rely daily on concepts and techniques I learned there. As a professor I regularly refer new students to my LSU course materials to explain the basics of light scattering and SEC.”

Virtual Method Optimization and Size Analysis for FFF

Flow-field flow fractionation (flow-FFF) offers highly versatile separations for the analysis of complex fluids, covering a size range of macromolecules and particles from 1 nm to 10,000 nm. However, flow-FFF is often perceived as a difficult technique to learn because of the multiple parameters available for adjustment.

Our new SCOUT DPS® software simulates flow-FFF from basic physical principles, enabling the virtual optimization of flow-FFF methods and opening up the power of flow-FFF separations to non-experts. SCOUT DPS also analyzes particle size distributions (hydrodynamic radius) by elution time to increase characterization capabilities. This publication in The Column explains how it works. http://www.chromatographyonline.com/field-flow-fractionation-virtual-optimization-versatile-separation-methods-0.

FFF Illustration

Wyatt Customer to Win Nobel Prize in Chemistry

Wyatt Technology congratulates its customer, Dr. Joachim Frank of Columbia University on sharing the Nobel Prize in Chemistry today with Jacques Dubochet and Richard Henderson. We are delighted to see their work developing cryo-electron microscopy recognized for its use in high-resolution structure determination of biomolecules in solution. Some of Professor Frank’s publications using our DAWN, Optilab and NanoStar instruments may be found in our Bibliography. Here is a recently published paper in Science;
Y. Chen, O. B. Clarke, J. Kim, S. Stowe, Y.-K. Kim, Z. Assur, M. Cavalier, R. Godoy-Ruiz, D. C. von Alpen, C. Manzini, W. S. Blaner, J. Frank, L. Quadro, D. J. Weber, et al.. Structure of the STRA6 receptor for retinol uptake. Science 2016 353(6302).

This is just a sample of the many peer-reviewed articles utilizing Wyatt instruments that may be found by searching our online Bibliography.

Nobel Prize

   

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Wyatt Technology is the recognized leader in light scattering instrumentation and software for determining the absolute molar mass, size, charge and interactions of macromolecules and nanoparticles in solution.

Wyatt's line of multi-angle static light scattering products couple to size exclusion chromatography (SEC-MALS), field-flow fractionation (FFF-MALS), and stop-flow composition-gradient systems (CG-MALS). Our dynamic light scattering (DLS) products operate in traditional cuvette as well as on-line and automated, high-throughput modes. We also offer unique instruments for electrophoretic light scattering (MP-PALS), differential refractometry, and differential viscosity.



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