Thermostability of self-assembled protein aggregates
(2022) KBKM01 20221Pure and Applied Biochemistry
- Abstract
- Nanoparticles used as drug delivery vehicles for small and large active drug molecules is a highly sought-after area of research. Nanoparticles made using recombinant proteins prove to be more beneficial as they are biocompatible and biodegradable, however only a few proteins can be considered as their availability and stability are not known. One such protein that has the potential to form nanoparticles which can be used as drug vehicles is known as amelogenin. It is a protein that is produced by cells known as ameloblasts during the amelogenesis process and helps in the formation of the enamel layer of teeth. Amelogenin has the ability to self-assemble into nanoparticles known as nanospheres and it a highly pH dependent process. In this... (More)
- Nanoparticles used as drug delivery vehicles for small and large active drug molecules is a highly sought-after area of research. Nanoparticles made using recombinant proteins prove to be more beneficial as they are biocompatible and biodegradable, however only a few proteins can be considered as their availability and stability are not known. One such protein that has the potential to form nanoparticles which can be used as drug vehicles is known as amelogenin. It is a protein that is produced by cells known as ameloblasts during the amelogenesis process and helps in the formation of the enamel layer of teeth. Amelogenin has the ability to self-assemble into nanoparticles known as nanospheres and it a highly pH dependent process. In this project, three variants of amelogenin were produced and purified. Amelogenin self-assembled into nanospheres using different buffers, which had an average hydrodynamic diameter of 33 to 40 nm. The produced nanospheres were then lyophilized and subjected to different temperatures and different durations to observe their stability and shelf-life. Finally, the structure of the formed amelogenin nanospheres were studied using structural and analytical techniques such as Mass Spectrometry and Fourier Transform Infrared spectroscopy (FTIR). Studying the self-assembling properties of amelogenin can give more insight on their use as drug carriers, which could have a major potential in the pharmaceutical and medical industry. (Less)
- Popular Abstract
- Nanoparticle drug delivery is the future!
This concept originally came about a century ago, where the famous scientist Paul Ehrlich talked about “magic bullets” that can target and kill harmful organisms in a body, without harming the host. This has prompted many scientists to find an effective way to deliver drugs to a particular site in the body, however there were problems that arose such as the potency of the drug once in the body and also how to direct the drug to the site of injury. This research has led to the discovery of nanoparticles or nanospheres which are very minute structures that can be made using biological and non-biological materials, on which the drugs can be attached and can finally enter the body.
In this... (More) - Nanoparticle drug delivery is the future!
This concept originally came about a century ago, where the famous scientist Paul Ehrlich talked about “magic bullets” that can target and kill harmful organisms in a body, without harming the host. This has prompted many scientists to find an effective way to deliver drugs to a particular site in the body, however there were problems that arose such as the potency of the drug once in the body and also how to direct the drug to the site of injury. This research has led to the discovery of nanoparticles or nanospheres which are very minute structures that can be made using biological and non-biological materials, on which the drugs can be attached and can finally enter the body.
In this study, a protein known as amelogenin was used to make nanospheres. Amelogenin is a protein that helps in the formation of tooth enamel, and has a unique property of assembling into small nanosized assemblies, such as nanospheres, at certain pH conditions. The study makes use of specially designed amelogenin proteins to test temperature stability of self-assembled protein nanospheres. This is important for their potential use in drug delivery.
The proteins were produced in bacteria and purified for further studies. Here, buffers of different salt contents and concentrations have been used to initiate the nanosphere formation. These buffers were added to the purified proteins, which were left for an hour for the nanospheres to form. After an hour, the sizes of the nanospheres formed were analyzed by a device that made use of a technique known as light scattering, where the sizes of the particles are determined based on how the light bounces off these particles. After noting down the sizes, the nanospheres were made into a solid form by a process known as freeze-drying, where frozen samples are kept in vacuum to remove all the water from the samples, making them completely solid and dry.
The freeze-dried samples, were then exposed to different temperatures such as room temperature, 37°C, 50°C and 70°C for one day. This is to study how the nanospheres would react to these different temperatures and to observe if there are any changes in their structure after they have been exposed to these temperatures.
Another temperature study was done for 1 week, where the nanospheres were exposed to a temperature of 37°C, after which their sizes were observed. This study was done to see how long the nanospheres could remain stable, which could translate to how long these nanospheres can last in a human body as our body temperature is around 37°C.
Finally, these samples were studied to detect any particular structural changes that could have occurred before and after heating the samples, and if there was any degradation of the proteins in the nanospheres because of the heat.
The stability of these nanospheres at different temperatures and time periods gives us an understanding of how long these nanospheres can be stored in the long run and how they may react if introduced into a human, for medical purposes. These results and conclusions pave a way in the pharmaceutical and medical industry in a way that has never been explored before! (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9084879
- author
- Mahesh, Divya LU
- supervisor
- organization
- course
- KBKM01 20221
- year
- 2022
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Amelogenin, Nanospheres, Temperature studies, Lyophilization, Biotechnology, Biochemistry
- language
- English
- id
- 9084879
- date added to LUP
- 2022-06-09 14:49:21
- date last changed
- 2022-06-09 14:49:21
@misc{9084879,
abstract = {{Nanoparticles used as drug delivery vehicles for small and large active drug molecules is a highly sought-after area of research. Nanoparticles made using recombinant proteins prove to be more beneficial as they are biocompatible and biodegradable, however only a few proteins can be considered as their availability and stability are not known. One such protein that has the potential to form nanoparticles which can be used as drug vehicles is known as amelogenin. It is a protein that is produced by cells known as ameloblasts during the amelogenesis process and helps in the formation of the enamel layer of teeth. Amelogenin has the ability to self-assemble into nanoparticles known as nanospheres and it a highly pH dependent process. In this project, three variants of amelogenin were produced and purified. Amelogenin self-assembled into nanospheres using different buffers, which had an average hydrodynamic diameter of 33 to 40 nm. The produced nanospheres were then lyophilized and subjected to different temperatures and different durations to observe their stability and shelf-life. Finally, the structure of the formed amelogenin nanospheres were studied using structural and analytical techniques such as Mass Spectrometry and Fourier Transform Infrared spectroscopy (FTIR). Studying the self-assembling properties of amelogenin can give more insight on their use as drug carriers, which could have a major potential in the pharmaceutical and medical industry.}},
author = {{Mahesh, Divya}},
language = {{eng}},
note = {{Student Paper}},
title = {{Thermostability of self-assembled protein aggregates}},
year = {{2022}},
}