Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

NIST develops NMR 'fingerprinting' for monoclonal antibodies

16.04.2015

National Institute of Standards and Technology (NIST) researchers at the Institute for Bioscience and Biotechnology Research (IBBR) have demonstrated the most precise method yet to measure the structural configuration of monoclonal antibodies (mAbs), an important factor in determining the safety and efficacy of these biomolecules as medicines.

Monoclonal antibodies are proteins manufactured in the laboratory that can target specific disease cells or antigens (proteins that trigger an immune reaction) for removal from the body. The method described in a recent paper* may soon help manufacturers and regulators better assess and compare the performance and quality of mAbs.


A schematic showing the NISTmAb monoclonal antibody, an immunoglobulin G (IgG) molecule being developed by NIST as a reference material. The labels mark the fragments Fab and Fc that were used in the novel NIST two-dimensional NMR fingerprinting method to measure the structural configuration of the entire antibody.

Credit: NIST

The IBBR is a joint institute of NIST and the University of Maryland.

Monoclonal antibodies can be used as extremely specific therapeutic agents, including ones designed to target cancer cells unique to an individual. However, in order to properly function as a biotherapeutic agent, the molecule's structural units--amino acids--must fold into a three-dimensional structure that aligns its active regions with corresponding receptor sites on a target cell or antigen.

If misfolding occurs, a potent and safe treatment may become ineffective, or worse, provoke a dangerous or fatal immune reaction. High-resolution spectral analysis--imaging at the atomic level where even the bonds between hydrogen and carbon atoms are distinguishable--is required to precisely define the mAb's structure and determine if the protein is folding properly.

"We refer to this as 'measuring fingerprints,' because just as a person has a unique set of fingerprint patterns, each mAb has a one-of-a-kind spectral makeup," says NIST research chemist Robert Brinson. "If we can map that spectral fingerprint, we can determine whether or not folding is occurring as desired."

To do this, the IBBR team turned to a solution that would surprise most biopharmaceutical experts: two-dimensional nuclear magnetic resonance (2D NMR) spectroscopy. NMR is a technique that measures the atomic signature of a molecule similar to how doctors use magnetic resonance imaging (MRI) to noninvasively view organs. "To date, it's been assumed that 2D NMR could not be practically applied to monoclonal antibodies because it's too insensitive, too time intensive and too expensive for analyzing anything other than much smaller drug molecules," Brinson explains.

In pushing the boundaries of the technique, the IBBR team used an NMR system with a high magnetic field strength to produce the first 2D NMR map of a complete, drug-like mAb.** The map was generated using signals from methyl groups.

"Methyl groups are dispersed throughout the mAb structure and, in particular, in the folded cores of the molecule that we want to evaluate," Brinson says. "We can use their signals to yield a specific spectral fingerprint that reflects the unique structure of the mAb."

To make the 2D NMR method more accessible to the lower-strength magnetic field instruments found in most analytical research labs, the IBBR team narrowed the analysis by dividing its sample antibody into two structural fragments."We mapped the 2D NMR signals generated by the subset of methyl groups found in these fragments, both about a third of the size of the entire protein," Brinson says. "The sum of the data gained from this analysis was found to be a good proxy for the spectral fingerprint of the full mAb."

The new 2D NMR fingerprinting method also overcomes the problems of cost and time. "We reduced the time needed for our measurements from many hours to about 30 minutes," Brinson says.

Brinson says that he and his colleagues are now working on a statistical method that will allow users of their 2D NMR methodology to compare fingerprints from multiple protein samples. "With that ability, manufacturers will be able to quantitatively show that spectra obtained from different lots of the same drug product are identical, enabling them to better meet regulatory requirements for quality and performance," he says.

###

* L.W. Abrogast, R.G. Brinson and J.P. Marino. Mapping monoclonal antibody structure by 2D 13C NMR at natural abundance. Analytical Chemistry, 87: 3556-3561 (2015). DOI: 10.1021/ac504804m

** The monoclonal antibody used in this experiment is NISTmAb, an immunoglobulin G type 1 donated by MedImmune and being developed by NIST as a reference material.

Media Contact

Michael E. Newman
michael.newman@nist.gov
301-975-3025

 @usnistgov

http://www.nist.gov 

Michael E. Newman | EurekAlert!

More articles from Life Sciences:

nachricht Biochemical 'fingerprints' reveal diabetes progression
22.08.2017 | Umea University

nachricht When fish swim in the holodeck
22.08.2017 | University of Vienna

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

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

Im Focus: Fizzy soda water could be key to clean manufacture of flat wonder material: Graphene

Whether you call it effervescent, fizzy, or sparkling, carbonated water is making a comeback as a beverage. Aside from quenching thirst, researchers at the University of Illinois at Urbana-Champaign have discovered a new use for these "bubbly" concoctions that will have major impact on the manufacturer of the world's thinnest, flattest, and one most useful materials -- graphene.

As graphene's popularity grows as an advanced "wonder" material, the speed and quality at which it can be manufactured will be paramount. With that in mind,...

Im Focus: Exotic quantum states made from light: Physicists create optical “wells” for a super-photon

Physicists at the University of Bonn have managed to create optical hollows and more complex patterns into which the light of a Bose-Einstein condensate flows. The creation of such highly low-loss structures for light is a prerequisite for complex light circuits, such as for quantum information processing for a new generation of computers. The researchers are now presenting their results in the journal Nature Photonics.

Light particles (photons) occur as tiny, indivisible portions. Many thousands of these light portions can be merged to form a single super-photon if they are...

Im Focus: Circular RNA linked to brain function

For the first time, scientists have shown that circular RNA is linked to brain function. When a RNA molecule called Cdr1as was deleted from the genome of mice, the animals had problems filtering out unnecessary information – like patients suffering from neuropsychiatric disorders.

While hundreds of circular RNAs (circRNAs) are abundant in mammalian brains, one big question has remained unanswered: What are they actually good for? In the...

Im Focus: RAVAN CubeSat measures Earth's outgoing energy

An experimental small satellite has successfully collected and delivered data on a key measurement for predicting changes in Earth's climate.

The Radiometer Assessment using Vertically Aligned Nanotubes (RAVAN) CubeSat was launched into low-Earth orbit on Nov. 11, 2016, in order to test new...

Im Focus: Scientists shine new light on the “other high temperature superconductor”

A study led by scientists of the Max Planck Institute for the Structure and Dynamics of Matter (MPSD) at the Center for Free-Electron Laser Science in Hamburg presents evidence of the coexistence of superconductivity and “charge-density-waves” in compounds of the poorly-studied family of bismuthates. This observation opens up new perspectives for a deeper understanding of the phenomenon of high-temperature superconductivity, a topic which is at the core of condensed matter research since more than 30 years. The paper by Nicoletti et al has been published in the PNAS.

Since the beginning of the 20th century, superconductivity had been observed in some metals at temperatures only a few degrees above the absolute zero (minus...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

Event News

Call for Papers – ICNFT 2018, 5th International Conference on New Forming Technology

16.08.2017 | Event News

Sustainability is the business model of tomorrow

04.08.2017 | Event News

Clash of Realities 2017: Registration now open. International Conference at TH Köln

26.07.2017 | Event News

 
Latest News

Molecular volume control

22.08.2017 | Life Sciences

When fish swim in the holodeck

22.08.2017 | Life Sciences

Biochemical 'fingerprints' reveal diabetes progression

22.08.2017 | Life Sciences

VideoLinks
B2B-VideoLinks
More VideoLinks >>>