The summary
was held after the experimental part was basically finished, mainly focusing on the
commercial outlook of our products. We had an in-depth communication with the professor who
is familiar with industrialization, and also had a meeting with relevant investment
institutions through the platform of Kingsray International Synthetic Biology & Gene & Cell
Therapy Global Industry Chain Forum. The following information will show the results of the
product commercialization outlook exchange.
Product
background
This year, we
further modified the chassis organisms Vibrio natriegens based on 2018 Murburg’s project to
develop our collection. V. natriegens is currently known as the fastest-growing
bacteria among all the chassis organisms. Under sufficient conditions, the generation time
is less than 10 minutes, and the growth rate is nearly twice as fast as that of the
traditional model organism Escherichia coli. V. natriegens possesses natural
competence, which can directly intake DNA from the environment, eliminating the process of
transformation. Therefore, the application of V. natriegens will save a lot of time
and reduce costs whether it is in scientific research or industrial production. At the same
time, V. natriegens has a wide range of substrate sources and it can express proteins
in high product yields using simple carbon sources and it is a novel model organism with
great potential to replace E. coli. We further characterized the expression elements
in V. natriegens and realize regulation of the gene circuit, providing a further
practical basis for V. natriegens as a novel model organism.
Current
situation of chassis biological applications
The current
chassis organisms used in various laboratories include Saccharomyces cerevisiae, B.
bischeri, E. coli, Bacillus subtilis, Bacillus glutamicus, Bacillus licheniformis,
Pseudomonas malodorosa, Lactobacillus, Methanophilus, Salmonella, Cyanobacteria,
Actinomyces, and some molds, etc. The widely used prokaryotic and eukaryotic chassis
organisms are E. coli and Saccharomyces cerevisiae respectively. The gene
expression elements associated with these two types of chassis organisms are well developed
and allow for precise and controlled gene regulation.
Limitations
and disadvantages of current chassis biology
1) Long
incubation time: for the two most common chassis organisms, S. cerevisiae typically
takes 48-96 h to form visible colonies in a constant temperature culture medium, Although
E. coli has a shorter incubation time, its culture time also reaches 12-16h.
Molecular cloning is the basis of many synthetic biology projects, and the culture process
for chassis organisms takes up most of the experimental time and greatly lengthening the
timeline of the experiment.
2) The
conversion process is time-consuming and the conversion rate is low: The transformation of
various chassis organisms is usually thermal, electrical, or chemical transformation, which
is time-consuming, and the conversion rate is affected by many factors, thus the conversion
process is unstable.
3) Microbial
culture medium is expensive: microbial culture medium usually needs to include water, carbon
sources, nitrogen sources, salts, and growth factors. Sufficient nutrients can provide the
necessary raw materials and sufficient energy for bacterial metabolism, growth, and
reproduction. However, various substrates also make the price of the culture medium becomes
expensive. For example, the price of MD medium used by S. cerevisiae is 480-980 CNY.
4) Slow
protein synthesis rate in chassis organisms: Ribosomes, as the organelles mainly responsible
for protein synthesis in bacteria, determine the rate of the synthesis. According to
statistics, E. coli can produce about 90,000 ribosomes per cell at a generation time
of 25 minutes, compared to about 115,000 for V. natriegens, which has a faster rate
of heterologous protein expression and greatly accelerates the step of molecular cloning.
The role of
chassis organisms in biopharmaceuticals
The
application of synthetic biology in biopharmaceuticals is the use of chassis organisms to
produce products with clinical value on a large scale. It starts from the most basic
elements and builds the parts step by step and relies on the chassis organisms for
regulation at the gene level and protein expression, etc., thus realizing artificially
designed genetic circuits that function as biosynthesis. The aim of synthetic biology is to
create artificial biosystems to regulate production processes and make them work like
circuits, and all this needs to be done in chassis organisms, such as E. coli, so the
characteristics of the chassis organisms and their modification play a crucial role in
biopharmaceuticals. For better engineering, the development of gene regulation and
expression elements in different chassis organisms is necessary to help us build a "cell
factory" using the different characteristics of the chassis organisms to improve the
efficiency of biosynthesis, etc.
Biopharmaceutical Market Development Status and Prospects
Figure 15.Global Biosimilars Market Size Statistics
The global
biosimilars market size has maintained a high growth rate between 2015 and 2019, and the
biosimilars market has reached a size of USD 9.5 billion in 2019, which has achieved more
than 3 times growth compared to 2015. It is evident that there is a strong demand for
biosimilar drugs in the market. And with the unstable outbreak of global epidemics, the
demand for vaccines has increased, so it can be predicted that the future biosimilar market
size will grow steadily with extensive prospects.
Figure 16.China Biosimilars Market Size Statistics
China's
biosimilar market has developed in recent years but the market size is still much smaller
than that of countries such as Europe and the United States since it starts late. However
its growth rate is on par with the global market, and as major enterprises resume work and
production after the epidemic, the production volume will gradually rise and the growth rate
will accelerate. In response to the rapid development of demand, the emerging market
represented by the Chinese market will gradually increase its market share in
biopharmaceuticals and enter a “golden period”.
The problem
faced by biosimilar production is the lack of efficient chassis organisms that have a high
expression level of heterogeneous protein and large-scale industrial production processes,
so the growing market has an urgent need for efficient chassis organisms, stable protein
expression systems, and the prospect of inexpensive chassis organisms to further improve
industrial-scale production.
Product
analysis (PEST analysis)
Politics: The
project "Design and Construction of Extreme Microbial Chassis Cells" of the National Key
Research and Development Program is promoting the research on microbial chassis cells in
extreme environments, establishing an efficient genome editing system for extreme
microorganisms, designing and synthesizing stable and universal heat-, salt-, alkali- and
acid-resistant biological device modules, researching and constructing extreme microbial
metabolic networks. The project aims to construct advanced versions of chassis cells to
adapt to specific environments and special growth conditions and carry out large-scale
industrial applications. It can be seen that the research on salt-resistent extreme
microbial chassis cells, such as V. natriegens, will develop rapidly under the
support of policies.
Economics:
There is a huge market demand for inexpensive, fast-growing chassis organisms that allow for
stable gene regulation. However, the corresponding chassis organism gene expression elements
are very scarce, poorly developed, and still unable to achieve the same level of gene
precision and stable protein expression as the commonly used chassis organisms such as E.
coli. Therefore, there are few of these fast-growing chassis organisms available. As can be
seen, there is a huge gap between supply and demand.
Society:
According to our social survey, the target population has a higher willingness to consume
more convenient, more affordable and faster growing sump organisms, reaching 85.98%, 80.37%,
and 75.7% respectively, which shows that can V. natriegens meet the rigid demand of
the target users.
Technology:
current gene expression elements such as promoters for V. natriegens based on E.
coli may not be well suited for use in V. natriegens. Meanwhile, 2018
Marburg's Colloection is convenient for assembly but may not be suitable for the regulation
of its gene expression. Therefore, our project developed and characterized more applicable
V. natriegens, and achieved the construction and expression regulation of gene
circuits in V. natriegens, providing a basis for practical applications in V.
natriegens.
