schedule – aiby 21 mai 2008
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schedule – AIBY 21 Mai 2008
08:30
• complementary competences (groups presentations)
• proposal of joint projects
• collaborators/competitors outside of AIBY
• arguments to EU 12:15
• vision
• AIBY focus (expression systems, products, technologies)
• next steps (schedule, To Do)
end 15:00
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resources
http://www.biotech2008.ch
aiby2009
http://cms.biotechlab.net/information/materials_aiby/
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Wädenswil
Bioprocess Technology
Institute of Biotechnology (IBT)
School of Life Sciences and Facility Management
Zurich University of Applied Sciences (ZHAW)
Waedenswil
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Zürich University of Applied Sciences
the largest multidisciplinary UAS, > 6000 students
8 departments (Wädenswil, Winterthur, Zurich)
Masters’ Programme in Pharmaceutical Biotechnology
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bioprocess technology
processtechnology
"software" forbiotechnology
high-productivity processesmicrobial glycosylation (E. coli, P. pastoris)
dynamics of physiologyfedbatch, flowcytometry
process control and simulation(software sensors, predictive control)
computer-intensive methodssignal extraction, clustering, etc.
eLearning www.biotechLAB.netknowledge managementadvanced training
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high-productivity processes
1
7
methanol:glucose mixtures – trypsinogen
5
4
3
2
1
0
CD
W
glu
cose
m
eth
ano
l (
g L
-1)
121086420
time (h)
5
4
3
2
1
0
met
abo
lite
form
iate
(g
L-1
)
2.5
2.0
1.5
1.0
0.5
0.0
pro
du
ct (
U L
-1)
A B DC E
8
HCD-fedbatch Mut+ P. pastoris – trypsinogen
120
100
80
60
40
20
0
CD
W
met
han
ol
(g
L-1
)
3020100-10-20
production time (h)
300
250
200
150
100
50
0
pro
du
ct (
U L
-1)
A B C
open symbols – 100% methanolclosed symbols – 40%:60% mixture of glucose and methanol
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10
product forms – Pichia pastoris
* product forms
rP productivity
rL rate of proteolytic degradation
rA rate of autocatalytic degradation
methanolbiomass (CDW)
metabolites
product 1product 2*product 3*...product i *
heat
O2
P. pastoris, Mut+
CO2
OH-/H+
methanol accumulation
proteases
rL
rA
rPdegradation product adegradation product bdegradation product c*...degradation product j *
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microbial glycosylation
2
12
US$ 5000 per mg – mGM-CSF
16
14
12
10
8
6
4
2
0
f
eed
rat
e (g
CH
3O
H L
-1 h
-1)
100806040200
time (h)
400
300
200
100
0
mG
M-C
SF
(m
g L
-1)
120
100
80
60
40
20
0
CD
W (
g L
-1)
13
yield and glycosylation pattern = f (physiology)
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yield and glycosylation pattern = f (physiology)
start of induction
end of cultivation
reference glycans
malto-dextrose reference
start of induction
end of cultivation
reference glycans
malto-dextrose reference
Man5GlcNAc2
15
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single cell analysis
3
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single cell analysis
averaged population subpopulations
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100
80
60
40
20
0
50403020100
production time (h)
100
80
60
40
20
0
PI
stai
ned
(%
)
10 %
porcine trypsinogen, pH 4, (methanol accumulation < 80 g L-1
, Figure 1) porcine trypsinogen, pH 4, stepwise decreasing methanol addition porcine trypsinogen, pH 5 horseradish peroxidase, pH 4 horseradish peroxidase, pH 5 wilde type X33 strain, pH 4
vitality by FCM (Mut+ P. pastoris)– single cell analysis
100
80
60
40
20
0
50403020100
production time (h)
100
80
60
40
20
0
PI
stai
ned
(%
)
10 %
porcine trypsinogen, pH 4, (methanol accumulation < 80 g L-1
, Figure 1) porcine trypsinogen, pH 4, stepwise decreasing methanol addition porcine trypsinogen, pH 5 horseradish peroxidase, pH 4 horseradish peroxidase, pH 5 wilde type X33 strain, pH 4
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complex functionalised biomolecules
1+2 = 4collagene III(high titre,
proper structure)
growth factor(e.g. PDGF )+ = dental implant
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complex functionalised biomolecules
1+3 = 5novel
promotorsphysiology+ = knowledge
/advice base
http://www.biotechLAB.net
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> 1000 proteins – Pichia pastoris
biomaterials(polymers)
pharmaceuticals
enzymes
peptides
http://faculty.kgi.edu/cregg/Pichia2004.htm
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5 + 1 high-value product categories – yeasts (Pichia pastoris)
• therapeutic proteins
• (catalytic proteins)
• enzymes for the synthesis/modification of pharmaceuticals
• new (functional) biomaterials for (e.g. health care)
• drug target proteins
• proteins for diagnostics
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