System Biology in downstream processing tech

System Biology in downstream processing

1. System Biology In Downstream
Processing
Vrushali Dengle

2. What is Systems Biology?????
The study of the mechanisms underlying
complex biological processes as integrated
systems of many interacting components.
involves
1.Collection of large sets of experimental data
2.proposal of mathematical models that might account for at least
some significant aspects of this data set
3.accurate computer solution of the mathematical equations to obtain
numerical predictions, and
4.assessment of the quality of the model by comparing numerical
simulations with experimental data

4. Mathematicians translate the results gained by cell
biologists, biochemists and microbiologists in their
laboratories into differential equations.
Bioinformaticians subsequently create computer
simulations.
System biologists are looking for the whole range of
natural processes and their constant interactions

6. Transcriptomics: whole cell or tissue gene
expression measurements by DNA microarraysor
SAGE
Proteomics:complete identification of proteins and
protein expression patterns of a cell or tissue
through two-dimensional gel electrophoresisand
mass spectrometryor multi-dimensional protein
identification techniques

7. Metabolomics:identification and measurement of
all small-molecules metaboliteswithin a cell or tissue
Interactomics:which is used mostly in the context
of protein-protein interaction but in theory
encompasses interactions between all molecules
within a cell
Fluxomics:,which deals with the dynamic changes
of molecules within a cell over time

8. What is the benefit of systems
biology?
for theoretical research, bioprocess and technical
applications as well as in biomedical applications.
“creating virtual cells” that reflect the functional
correlations of the individual cell components –also
on the molecular level –and simulate reactions to
external influences might lead to a holistic
understanding of the life functions of living things
and mechanisms that lead to diseases

9. Impact of system biology for the
improvement of wine yeasts and wine
making practices

10. System biology in down stream
processing

11. MBP fusion protein
In the pMAL™ Protein Fusion and Purification
System, the cloned gene is inserted into a pMAL
vector down-stream from themalEgene, which
encodes maltose-binding protein (MBP). This
results inthe expression of an MBP-fusion
protein.
The technique uses the strong Ptac promoter and
the translation initiation signals of MBP to express
large amounts of the fusion protein. The fusion
protein is then purified by a one-step affinity
purification specific for MBP

12. Reliable expression: substantial yields (up to 100 mg/L) in more
than 75% of the cases tested
Expression in either the cytoplasm or periplasm: periplasmic
expression enhances folding of proteins with disulfide bonds
Fusion to MBP has been shown to enhance the solubility of
proteins expressed in E. coli
Gentle elution with maltose: no detergents or harsh denaturants

14. ‘His’ tagged proteins-IMAC
Hisbinds with good selectivity to matrices incorporating Ni+2
ions, typically immobilized with either iminodiacetic acidor
nitrilotriacetic acidchelating groups.
The technique is known as immobilized metal affinity
chromatography.
The first steprelies on affinity purification of the N-terminal
His-tagged protein in the conventional way, eluting the protein
with imidazole.
Second step:Addition of a His-tagged 3C protease to cleave
the His-tag permits a second pass through the nickel column,
this time all impurities bind to the column while the pure protein
does not.

15. Eg: the application of this system to over-express and purify ORF
PA5104 from Pseudomonas aeruginosa. The protein was
successfully crystallized and crystals were shown to diffract to
atomic resolution.

17. GST tagged proteins
GST-tagged proteins bind noncovalentlyto an
affinity matrix composed of glutathione immobilized
on a chromatographic support.
The tag is displaced and the protein eluted with
solutions of glutathione.
GST can be used alone or in combination with His
tags to provide an alternate affinity purification
regime when IMAC conditions are not suitable.

18. Computational systems biologyis the algorithm and application
development arm of systems biology. It is also directly associated with
bioinformaticsand computational biology.
Computational systems biology aims to develop and use efficient
algorithms, data structuresand communication tools to orchestrate the
integration of large quantities of biological data with the goal of
modelling dynamic characteristics of a biological system.
The goal is to create accurate real-time models of a system's
response to environmental and internal stimuli, such as a model
of a cancer cell in order to find weaknesses in its signaling
pathways, or modeling of ion channel mutations to see effects
on cardiomyocytes and in turn, the function of a beating heart

19. The Future of Systems Biology
can be used to cut R&D time and costs through improved target
identification and lead quality, better elucidation of ADME/Tox profiles, as
well as optimizing clinical trial efficiency.