Evaluation of a novel drug depletion device for removal of biological drugs from human serum
(2016) MOBM01 20161Degree Projects in Molecular Biology
- Abstract
- Biological drugs such as tumor necrosis factor (TNFα) blockers are very successful and effective in therapy of chronic inflammatory disease. These blockers bind TNFα and inhibit its pro-inflammatory function. But unfortunately, biological drugs are immunogenic and prolonged treatment triggers the formation of anti-drug antibodies and neutralizing antibodies (NAbs). This reaction is associated with reduced clinical response and adverse effects. Therefore, it is of interest to detect the neutralization early during drug development and therapy. The iLite cell based assays has low tolerance for residual drug. NAbs can only be detected if they are higher in quantity than the drug. This means the assay underestimates the NAb concentration.... (More)
- Biological drugs such as tumor necrosis factor (TNFα) blockers are very successful and effective in therapy of chronic inflammatory disease. These blockers bind TNFα and inhibit its pro-inflammatory function. But unfortunately, biological drugs are immunogenic and prolonged treatment triggers the formation of anti-drug antibodies and neutralizing antibodies (NAbs). This reaction is associated with reduced clinical response and adverse effects. Therefore, it is of interest to detect the neutralization early during drug development and therapy. The iLite cell based assays has low tolerance for residual drug. NAbs can only be detected if they are higher in quantity than the drug. This means the assay underestimates the NAb concentration. Removing the drug specifically from the sample and release the NAbs from the complex would allow an earlier detection. In this project, newly developed gold nanoparticles coated with TNFα were tested as a drug depletion device. The nanoparticles were evaluated for their effectiveness in removing the biological drug infliximab (a TNFα-blocker) from a phosphate buffer model solution. Subsequently, the iLite cell based reporter gene assay was used for infliximab quantification or neutralizing antibody determination. Initial incompatibilities of the gold nanoparticles solution and the cell based assay were encountered, as the nanoparticle buffer solution distorted the assay signal. Consequently, the nanoparticles were optimized and the first step in depleting a simple model infliximab solution was successful. No infliximab was detectable after depletion. A subsequent depletion of infliximab from an infliximab-NAb complex model solution succeeded as well, but has potential to be improved. Future experiments have to be performed on infliximab-NAb complex spiked serum, on actual patient samples and for other TNFα-blockers. (Less)
- Popular Abstract
- Keep your eye on biological drugs!
Biological drugs have revolutionized the treatment of chronic inflammatory disease. In contrast to synthesized traditional small molecule drugs, biological drugs are much larger and naturally occurring structures. They are proteins, not chemicals. But unfortunately because of the size and protein-content, the drug is prone to trigger the immune system. This results in clearance or neutralization of the drug. If this happens, the patient will not respond to therapy and the symptoms of the disease will return. Continuing this highly costly treatment will not have any benefit for the patient. Therefore monitoring the treatment and spot the neutralization in an early stage is crucial.
Diagnostic tests... (More) - Keep your eye on biological drugs!
Biological drugs have revolutionized the treatment of chronic inflammatory disease. In contrast to synthesized traditional small molecule drugs, biological drugs are much larger and naturally occurring structures. They are proteins, not chemicals. But unfortunately because of the size and protein-content, the drug is prone to trigger the immune system. This results in clearance or neutralization of the drug. If this happens, the patient will not respond to therapy and the symptoms of the disease will return. Continuing this highly costly treatment will not have any benefit for the patient. Therefore monitoring the treatment and spot the neutralization in an early stage is crucial.
Diagnostic tests for therapy monitoring are already available on the market. The good part is that the test can detect anti-drug activity. The bad part is that the test gets disturbed by residual drug. The neutralization is only detectable if the anti-drug agent is higher in quantity compared to the drug. Thus removing the drug from the sample would give a better picture of the anti-drug activity state.
But first, why does the drug interfere with the test? This is because the drug forms complexes with the neutralization agent. If the agent is trapped in a complex, it cannot be uncovered. Therefore removing the drug from the sample would break the complex and free the neutralization agent for detection.
Hence in this project a drug removal step with nanoparticles was developed and evaluated.
Nanoparticles as drug catchers
These nanoparticles were developed to capture specifically a drug type or drug group and by subsequent centrifugation the pharmaceutical can be removed from the solution. This tool was very effective and specific. As shown in the bar graph, there was no drug left, when nanoparticles were applied on a drug solution. This means the drug was successfully removed.
The optimization of the test with a first drug depletion step would make possible to detect the neutralizing agent at very low concentrations. At which point the patient does not yet show signs of treatment failure; so the treatment can be adapted on time. It would avoid treatment with a non-effective drug and prevent the occurrence of adverse effects. Early detection has clinically and economically implications as the treatment with biological drugs is very expensive.
Supervisor: Lisbeth Witt, R&D Euro-Diagnostica, Malmö
Master’s Degree Project in Molecular Biology, 30 ECTS, 2016
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/8891706
- author
- Lang, Katharina
- supervisor
- organization
- course
- MOBM01 20161
- year
- 2016
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 8891706
- date added to LUP
- 2016-09-14 11:51:16
- date last changed
- 2016-09-14 11:51:16
@misc{8891706,
abstract = {{Biological drugs such as tumor necrosis factor (TNFα) blockers are very successful and effective in therapy of chronic inflammatory disease. These blockers bind TNFα and inhibit its pro-inflammatory function. But unfortunately, biological drugs are immunogenic and prolonged treatment triggers the formation of anti-drug antibodies and neutralizing antibodies (NAbs). This reaction is associated with reduced clinical response and adverse effects. Therefore, it is of interest to detect the neutralization early during drug development and therapy. The iLite cell based assays has low tolerance for residual drug. NAbs can only be detected if they are higher in quantity than the drug. This means the assay underestimates the NAb concentration. Removing the drug specifically from the sample and release the NAbs from the complex would allow an earlier detection. In this project, newly developed gold nanoparticles coated with TNFα were tested as a drug depletion device. The nanoparticles were evaluated for their effectiveness in removing the biological drug infliximab (a TNFα-blocker) from a phosphate buffer model solution. Subsequently, the iLite cell based reporter gene assay was used for infliximab quantification or neutralizing antibody determination. Initial incompatibilities of the gold nanoparticles solution and the cell based assay were encountered, as the nanoparticle buffer solution distorted the assay signal. Consequently, the nanoparticles were optimized and the first step in depleting a simple model infliximab solution was successful. No infliximab was detectable after depletion. A subsequent depletion of infliximab from an infliximab-NAb complex model solution succeeded as well, but has potential to be improved. Future experiments have to be performed on infliximab-NAb complex spiked serum, on actual patient samples and for other TNFα-blockers.}},
author = {{Lang, Katharina}},
language = {{eng}},
note = {{Student Paper}},
title = {{Evaluation of a novel drug depletion device for removal of biological drugs from human serum}},
year = {{2016}},
}