Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:


Neutron beams reveal how antibodies cluster in solution


Results from neutron spin-echo analysis at the Institut Laue-Langevin (ILL) and the National Center of Neutron Research (NCNR) in the United States are an important advance towards enabling subcutaneous injections of concentrated biopharmaceuticals used to treat cancer and autoimmune disorders (e.g. arthritis, multiple sclerosis). The insights obtained could help drug companies reduce the viscosity and mitigate phase separation in injectable biopharmaceuticals, making them easier to manufacture and fluid enough to be self-administered in the home.

Scientists have used small-angle neutron scattering (SANS) and neutron spin-echo (NSE) techniques for the first time to understand how monoclonal antibodies (mAbs), a class of targeted biopharmaceuticals used to treat autoimmune disorders and cancer, dynamically cluster and move in high concentration solutions. Certain mAb cluster arrangements can thicken pharmaceutical solutions; they could thus limit the feasible concentration of injectables administered to patients around the world.

© psdesign1 -

The insights provided by a team of neutron scientists from the National Center of Neutron Research (NCNR) and the Institut Laue-Langevin (ILL), in collaboration with colloid and proteins scientists at the University of Delaware and biopharmaceutical company Genentech (a member of the Roche group), are an important step towards the development and manufacture of high-concentration biopharmaceuticals needed for high-dose indications and potential self-administration at home.

Monoclonal antibodies (mAbs) are proving to be a vital tool in modern pharmacology, providing the basis for a growing number of successful drugs for cancer and autoimmune disorders such as arthritis and multiple sclerosis.  As agents for targeted therapy with a good safety profile, they are an alternative to harsher chemotherapy treatment.

The mAbs work by attaching themselves to specific protein targets on cancerous cells, or blocking target proteins in a known biochemical pathway responsible for a disease.  These treatments usually require high doses, and lately in some indications there has been considerable interest in moving from intravenous (IV) delivery to a more convenient subcutaneous (SC) delivery - a shallow injection into the cutis just below the skin (such as the home treatments offered to sufferers of type 1 diabetes). 

However, progress has been hampered by the high viscosity of solutions containing high amounts of mAbs, and this provides challenges to efficient and economical large-scale production, purification, and delivery of these drugs.

"For some proteins at concentrations higher than 100 mg/ml, you can’t deliver them fast enough through thin, SC injection needles, so repeat visits to the hospital with intravenous drips are needed," explains Prof. Yun Liu from the National Center of Neutron Research (NCNR) in the US, who is also affiliated with the University of Delaware. "The thickening may also cause problems in processing, when filtration pressures are too high, for example, or during freeze and thaw in large tanks where potential gelation or phase separation of the freeze concentrate can occur." 

As a result, efforts to find ways to raise the concentration of monoclonal antibody pharmaceuticals are focused on understanding the root cause of this thickening. Previous studies using static light scattering (Lilyestrom et al, 2013) on concentrated mAb solutions had suggested a strong link between the development of protein clustering formations and increases in viscosity.

In this latest study (Yearley et al. 2014)  led by Prof. Yun Liu (NCNR) and Dr. Dan Zarraga from Genentech, in collaboration with the Institut Laue-Langevin (ILL) in Grenoble, small-angle neutron scattering (SANS) and neutron spin-echo (NSE) techniques were used to study the structure and dynamics of mAb clustering responsible for the bulk solution properties. Two types of antibody were placed in solution - one known to increase solution viscosity and one which did not - so any differences in behaviour could be observed. 

Neutron spin-echo (NSE) measures cluster dynamics by determining their self diffusivity (as opposed to collective diffusivity measured in dynamic light scattering).  Neutrons are able to probe very high concentrations since they are scattered only by nuclei (these occupy very little space and thus appear dilute in the neutron beam’s perspective).  Also the unrivalled high resolution and very high neutron intensity provided by the ILL neutron spin-echo instrument IN15 allowed a systematic exploration of many different mAb samples.

Using this technique for the high viscosity mAb solution the team confirmed the presence of small extended clusters of mAbs with lifetimes sufficiently long to have an impact on bulk solution properties.  On the other hand, the diffusivity measured for the low viscosity mAb solution indicated no such clustering at timescales greater than 50 nanoseconds.  The two mAbs only differed in the sequence of their complementarity determining region (CDR); this shows that seemingly small changes in the sequence can have profound consequences on mAb solution behaviour.

Taking into account these insights on mAb clustering, recent studies (Zarraga et al 2013; Allmendinger et al 2014) have shown that there is a window of expulsion rates through a thin needle that reversibly disrupts these mAb clusters, helping mitigate the viscosity issue without irreversibly damaging the individual mAbs.  This provides a basis for designing an optimal device system for delivering biopharmaceutical injectable solutions at very high concentrations.

The progress made in these studies is due in part to bringing research institutions and industry together to address practical problems.  Research institutions in academe and national labs are often in search of applications for their technological solutions, whilst industry encounters practical problems when looking for solutions.  In addition, such collaboration can spur research in adjacent, often very important, fields.

Dr Peter Falus, instrument scientist at ILL said: "Whilst the potential impact of these studies on drug design is very exciting, the subject of protein clustering is an extremely interesting area in its own right. A lot of well-known phenomena, such as the cataracts in our eyes, or Alzheimer’s disease, are the results of proteins clustering in our bodies. As a physicist, I am interested in clustering in general, and neutron techniques here at the ILL provide a unique, high-resolution tool to investigate these complex interactions in natural organic systems."

For more information please contact James Romero  / Tel: +44 08456801866


1. Observation of Small Cluster Formation in Concentrated Monoclonal Antibody Solutions and Its Implications to Solution Viscosity, Yearley E et al, Biophys J 1763-1770 (15.04.2014)

2. Small Angle Neutron Scattering of mAb Conformations and Interactions at High Concentration, Yearley E et al, Biophys J 105:720-731 (2013).
3. Monoclonal antibody self-association, cluster formation, and rheology at high concentrations, Lilyestrom et al, J Phys Chem B 117:6373-84 (2013).
4. High shear rheology and anisotropy in concentrated solutions of monoclonal antibodies, Zarraga IE, Taing R, et al , J Pharm Sci  102: 2538–2549 (2013).
5. Rheological characterization and injection forces of concentrated protein formulations: An alternative predictive model for non-Newtonian solutions, Allmendinger A, et al Eur J Pharm Biopharm pii: S0939-6411 (Feb 18 2014)

Notes to editors


About ILL – The Institut Laue-Langevin (ILL) is an international research centre based in Grenoble, France. It has led the world in neutron-scattering science and technology for more than 40 years, since experiments began in 1972. ILL operates one of the most intense neutron sources in the world, feeding beams of neutrons to a suite of 40 high-performance instruments that are constantly upgraded. Each year 1,200 scientists from more than 30 countries worldwide visit ILL (2000 visits), to conduct research into condensed matter physics, (green) chemistry, biology, nuclear physics, and materials science. The UK, along with France and Germany, is an Associate and major funder of the ILL.

James Romero | idw - Informationsdienst Wissenschaft
Further information:

More articles from Life Sciences:

nachricht Molecular trigger for Cerebral Cavernous Malformation identified
26.11.2015 | EMBO - excellence in life sciences

nachricht Peering into cell structures where neurodiseases emerge
26.11.2015 | University of Delaware

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: Climate study finds evidence of global shift in the 1980s

Planet Earth experienced a global climate shift in the late 1980s on an unprecedented scale, fuelled by anthropogenic warming and a volcanic eruption, according to new research published this week.

Scientists say that a major step change, or ‘regime shift’, in the Earth’s biophysical systems, from the upper atmosphere to the depths of the ocean and from...

Im Focus: Innovative Photovoltaics – from the Lab to the Façade

Fraunhofer ISE Demonstrates New Cell and Module Technologies on its Outer Building Façade

The Fraunhofer Institute for Solar Energy Systems ISE has installed 70 photovoltaic modules on the outer façade of one of its lab buildings. The modules were...

Im Focus: Lactate for Brain Energy

Nerve cells cover their high energy demand with glucose and lactate. Scientists of the University of Zurich now provide new support for this. They show for the first time in the intact mouse brain evidence for an exchange of lactate between different brain cells. With this study they were able to confirm a 20-year old hypothesis.

In comparison to other organs, the human brain has the highest energy requirements. The supply of energy for nerve cells and the particular role of lactic acid...

Im Focus: Laser process simulation available as app for first time

In laser material processing, the simulation of processes has made great strides over the past few years. Today, the software can predict relatively well what will happen on the workpiece. Unfortunately, it is also highly complex and requires a lot of computing time. Thanks to clever simplification, experts from Fraunhofer ILT are now able to offer the first-ever simulation software that calculates processes in real time and also runs on tablet computers and smartphones. The fast software enables users to do without expensive experiments and to find optimum process parameters even more effectively.

Before now, the reliable simulation of laser processes was a job for experts. Armed with sophisticated software packages and after many hours on computer...

Im Focus: Quantum Simulation: A Better Understanding of Magnetism

Heidelberg physicists use ultracold atoms to imitate the behaviour of electrons in a solid

Researchers at Heidelberg University have devised a new way to study the phenomenon of magnetism. Using ultracold atoms at near absolute zero, they prepared a...

All Focus news of the innovation-report >>>



Event News

Fraunhofer’s Urban Futures Conference: 2 days in the city of the future

25.11.2015 | Event News

Gluten oder nicht Gluten? Überempfindlichkeit auf Weizen kann unterschiedliche Ursachen haben

17.11.2015 | Event News

Art Collection Deutsche Börse zeigt Ausstellung „Traces of Disorder“

21.10.2015 | Event News

Latest News

Using sphere packing models to explain the structure of forests

26.11.2015 | Ecology, The Environment and Conservation

Dimensionality transition in a newly created material

26.11.2015 | Materials Sciences

Revealing glacier flow with animated satellite images

26.11.2015 | Earth Sciences

More VideoLinks >>>