Structural Conformations of Monoclonal Antibodies at Oil/Water Interfaces

Lead Research Organisation: University of Manchester
Department Name: Physics and Astronomy

Abstract

Antibodies are proteins that bind to specific targets, helping the immune system fight infections and foreign substances. Monoclonal Antibodies (mAbs) are important in many biotechnology applications and are increasingly important in medicine, with treatments like immunotherapy and antibody chemotherapy resulting in a market worth over $100 billion.

Drugs containing mAbs or other proteins are often stored in pre-filled syringes to allow easier and less risky application. The barrel of these syringes is coated with oil, to allow the plunger to move without jamming. However, mAbs will often adsorb (stick) to the interface between oil and solution, which can disrupt their structure and thus their functionality. The presence of the oil/water interface can also cause disrupted proteins to form together into aggregates that can cause unwanted immune reactions in patients. To determine the full effect of the oil on the protein, and to find ways to minimise this effect, it is important to be able to determine the structure of proteins at these oil/water interfaces.

Currently, the investigation of proteins at oil/water interfaces is difficult, as the technique must be able to distinguish nanometre-scale structures through both solution and oil. Neutron reflection is a technique which is sensitive to nanometre scales, but until recently work at the oil-water interface was extremely difficult, as the beam of neutrons must pass through either a thick oil or solution layer, both of which will heavily attenuate the beam.

Prior to this project, work by M. Campana showed that this can be overcome by spin-coating the block to produce a thin oil layer, and deuterating the oil (replacing hydrogen in the oil with deuterium) to reduce the attenuation of the layer. However, this technique currently uses hexadecane, which is toxic but easy to deuterate, as a model oil. The method had also not yet been shown to work with mAbs, only reference proteins. Thus this project's goal is to develop the oil-water neutron reflection technique, along with other techniques like ellipsometry, to allow accurate investigation of mAbs at oil/water interfaces, and to investigate different the behaviour of different mAbs at the interface.

So far this project has shown that the hexadecane method also works with mAbs, giving a useful model system for mAbs at the oil/water interface. The focus of the project is now moving to measuring mAb adsorption to a more biologically compatible oil.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/N509565/1 01/10/2016 30/09/2021
1735875 Studentship EP/N509565/1 20/09/2016 30/09/2020 Sean Ruane
 
Description Antibodies are a part of the bodies immune system, genetically modified antibodies are used in immunotherapy and targeted drug delivery, especially in oncology. Antibody drugs are stored in pre-filled syringes, but they can adsorb to the oils used to coat these syringes, denaturing the antibodies and lowering the potency of the drug. So far we have proved that neutron reflection, a powerful technique for study very thin films, can be used to directly measure the conformation of antibodies at the oil-water interface, providing a powerful new tool to investigate the cause of antibody denaturation, and in developing tools to combat it.
Exploitation Route Further development of the technique is ongoing, as well as results looking at more complex antibodies, but the technique could be used to examine why specific drugs denature at the interface, or examine why certain processes, such as adding non-ionic surfactants, reduce denaturation, and how we can improve these techniques.
Sectors Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology