Team:TU Darmstadt/Human Practices/Expert Interviews
Expert Interviews
After getting a lot of input from various people of all fields and ages we decided to approach some experts in life sciences to elaborate further about the questions that came up. We are delighted that so many scientists decided to help us improve our project by sharing their knowledge with us. Down below you can find a short overview of the input every expert gave us. If you want to find out more about the talks we had with our experts please have a look at our big report.
Besides developing the scientific side of our project, we also
wanted to explore the impact our work has on society. After
talking with people of different ages and levels of education we
decided it would be best to contact an expert of both
ethics and
synthetic biology. Fortunately, we found
Prof. Sibylle Gaisser of the University of Ansbach who was
excited to help us, since she was already familiar with iGEM.
In our conversation she cautioned us about the
modularity
of our system, as its dual-use aspect might lead to
misappropriation. She emphasized the need for scientists to
consider the possibility, that their work might be used for
applications they did not intend it for.
Regarding our project she stressed the need to find out about
the immunological
reactions of the
human
body to our Virus-like particles (VLPs), regardless of the
application the particles are meant for.
Considering the location of our university she encouraged us to
contact the Paul-Ehrlich-Institute in
Langen, to ask whether they could help us with testing our VLPs.
For the entire conversation please have a look at our big report.
We encountered Dr. Jörg Mampel from the BRAIN AG during an event where he held a presentation at our university. After telling him about our project he offered us his help since he had previously worked with biological nanoparticles, so called Biomolecular Compartments (BMCs). The development of those was similar to that of our VLPs. Therefore, we spoke mostly about feasible purification and analyzation methods. He recommended us to use the electron microscope for analyzing our particles and shared some of his knowledge about industrial-scale production of agents. He also reassured us that our particles would not enclose foreign matter during assembly but nevertheless he stressed how important it is to test our particles for pyrogenic and foreign material. A very valuable information Dr. Mampel gave to us was the fact that he worked with different promoters that are induced by Isopropyl‑ β‑d‑1‑thiogalactopyranoside (IPTG) like the T7 promoter but found out that expression levels can be regulated better by using promoters induced by arabinose like the pBAD promoter. This was something we then implemented into our project having pDESTara2 as one of our vectors. If you want to read the entire interview please view our big report.
As one of our possible applications are vaccines, we were
interested in the challenges and requirements of producing
VLPs for medical applications. To inquire further about this
topic, we were happy to have the opportunity to talk with Dr. Jakob Cramer who is the
head of clinical development at the
Coalition for Epidemic Preparedness Innovations (CEPI) in
London. Asking him about the steps we would need to take if we
wanted to produce our VLPs for an application in the human body
he told us that checking on the immunogenicity and
toxicity is
the main point. He thinks that
our "real MVPs" pose an opportunity as a platform for mixed
vaccines especially because VLPs can serve as natural adjuvants.
If you want to read the complete interview please have a look at
our big report.
Picture 1: Source: DKFZ T. Schwerdt
Based on zur Hausen’s discovery of the role human papillomavirus (HPV) plays in the development of cervical cancer the development of standardized prophylactic vaccines against the cancerous and most common HPV strains was possible. These vaccines use a VLP designed and produced through recombinant technology. A possible application of our modular platform is the development of vaccines. Vaccines using VLPs may be safer, given the lack of DNA and therefore lack of infectiousness. The development of vaccines is a long process we hope could be shortened by the availability of our expression system and enable the addition of desired antibodies in as little as one cloning step. Through our conversation with Professor zur Hausen we hoped to gain insight in the process of the development of a vaccine, the communication of information about vaccines to the public, his thoughts on our project and his experiences in the scientific community. Prof zur Hausen spoke about vaccine-costs being high due to the long development and was optimistic about our platform lowering the costs of vaccine-development through shortening the developmental phase. Further we spoke about the government’s role in educating the public on vaccines, the effect media can have on conveying science and securing funding for research project. To read more, follow the link to our HP Report.
Wanting to focus on the industrial
production and hoping to gain
more insight about VLPs from experts working with them we called
the Fraunhofer Institute in
Stuttgart asking for Prof. Bailer.
She worked with VLPs aiming to develop new vaccines before.
Having a lot of knowledge about the benefits and downside of
VLPs she compared our project to their way of working. She told
us that modifying the particles after the assembly might pose a
big advantage because the coat protein will not be sterically
hindered by the fusion protein attached to it. Also, she was
enthusiastic about varying the
level of modification as they have not tried
it in their project yet. Nevertheless, she stressed that we
might need a lot of purification steps for our VLPs since we are
producing them in E. coli, which is an organism used in
production of medicine but also known for producing endotoxins.
She suggested to use ultracentrifugation to analyze the VLPs and
an electron microscope to take a look at the shape and size and
check the integrity. She also pointed out that it might be
possible that the particles lose their homogeneity if they are
modified. An aspect she stressed was to check for the
immunogenicity and toxicity of our particles
which is why she recommended us to visit the
Paul-Ehrlich-Institut, Germany. If you want
to know more about our conversation, please take a look at our
big report.
Getting a lot of feedback about toxicity and immunogenicity we
then approached Dr. Stefan Schülke to learn more about the
properties of our VLPs. As an expert for immunology he told us
that our VLPs should not be immunogenic without modification
since they do not contain viral DNA. This gives the opportunity
to use the VLPs for various applications in the human body.
Another big benefit he sees in our VLPs is their
modularity
. In his opinion our system could speed up the development of
vaccinations which would help to react faster to outbreaks or
newly upcoming diseases. Having the
P22 VLPs as a platform could help to
prepare for various diseases through passive immunization and
poses the benefit of easily producible mixed vaccines.
Furthermore, he told us that VLPs in general have a history of
safe use and that some, for example the HPV vaccine, are already
approved by authorities.
Additionally, we want to thank him for giving us the opportunity
to perform experiments on our VLPs at the
Paul-Ehrlich-Institut
(PEI) in Langen.
For the whole conversation please take a look at our
big report.
We are grateful for the fruitful discussion with Prof. Santi
about our project. Being an expert for the recombinant
expression of VLPs in plants he talked with us about other
expression systems than E. coli and
various methods for purification. Since we are interested in
an outlook for our system and the possible
applications, we further elaborated purification methods. He
recommended to use size exclusion if necessary, to produce our
particles in a larger scale. Regarding the different
applications he suggested to include a method for varying modification levels. On one hand this would pose the advantage of
particles not
being disturbed by steric hindrance during the assembly, and on
the other hand this would enable one to set the
level of modification fitting the desired
application. Vaccines for example might require less proteins
displayed on the outside than particles meant for drug delivery.
Our method of expressing coat proteins with and without
recognition tag for the sortase might - in
his opinion - be a solution for this problem.
For the entire conversation please take a look at our
big report.
Prof. Santi pointed out another expert on VLPs, Dr. Chiara Lico.
Together with her we had a helpful conversation about the
large-scale
production of VLPs and their
benefits and downsides. She told us that a major advantage is
the way of production because due to their self-assembly VLPs
can be produced easily and economically even at large scale.
Another benefit is the fact that VLPs of different classes vary
greatly in shape and dimension which gives the opportunity to
use them for many applications. They can be modified chemically
and genetically in various ways and thus shape the inner and
outer structure. In order to increase the capacity of our
purification methods, she recommended us to try
size exclusion chromatography. Also, she
suggested us to analyze the particles by an ELISA or a silver
stained SDS-PAGE. If we wanted to take a closer look at our
modified particles, she confirmed the best way would be to use
an electron microscope.
For the entire interview please take a look at our
big report.
