A glimpse into what we can offer at Signalchem in custom projects and profiling solutions

1. Custom Projects and Profiling

2. PAOLO LOBO
TECHNICAL PROJECT MANAGER

3. PROTEIN PRODUCTION PROCESS
Protein
Plasmid/
Virus
Expression vectors
Expression Evaluation
& Optimisation
Bacterial/Insect/Plant/
Mammalian cell culture
Protein
Purification
HPLC, Ion Exchange,
Gel filtration, Affinity
Chromatography
QC
SDS-PAGE, WB,
RP-HPLC, MS,
Activity assays
Gene Synthesis
& Subcloning
Primer design
Sequencing
Cloning
Package &
Delivery
Storage
Lyophilization
Shipping
PhD Scientists lead every department involved in Manufacture/QC Processes

4. PROTEIN PRODUCTS
Reagents Mostly assay reagents
(buffer, substrates, co-factors)
Active enzymes Kinases
Phosphatases
Phosphodiesterases (PDEs)
GTPases and ATPases
Ubiquitin enzymes
Epigenetic enzymes
Proteases
Proteins Adaptor proteins
Transcription proteins
Tau and phospho-Tau
Enzyme substrates

5. PRODUCT APPLICATIONS
• Screening
• Selectivity screens
• Inhibition profiles
• Kinetics
• Western blot
• As positive control
• Check if the protein is full-length
• Specific phosphorylation sites
• Crystallography
• Usually require much higher
concentration and purity
• Would prefer tag-free or
small tag (His but not GST)
• Binding studies
• ITC
• Thermodynamics
• Can be done in assay format
(screening)
• Custom applications
• Other research
• Open to a variety of suggestions
• Flexible
• Communication

6. CUSTOMIZATION OPTIONS
• Concentrating
• Buffer exchange
• Purity improvement
• Tag removal
• (De)phosphorylation
• Endotoxin removal
• Sterilization
• Lyophilization
• Fusion proteins
• More
Design
Feasibility Pricing QC
Turnaround
Custom Projects

7. EXPRESSION SYSTEM OFFERINGS
E. coli
Bacteria
1. “work horse”
2. Well established
3. High Expression
4. Simple Genetics
5. Easy Scale up
6. Speed
7. Low Cost
8. Equipment
S. Cerevisieae
P. Pastoris
Yeast
1. PTMs
2. Soluble Proteins
3. High Expressor
4. Easy Scale up
Tobacco
Plant
1. PTMs
2. Soluble Proteins
3. Easy Growing
4. Low Cost
5. Renewable
Sf9, Sf21, High 5
Insect
1. PTMs
2. Soluble Proteins
3. High Expressor
CHO, HEK, COS
Mammalian
1. PTMs
2. Soluble Proteins
3. Lower expression
4. Expensive

8. PROFILING SERVICES
Kinase
Compound concentration
Compounds
+
+
+
+
• Compound Selectivity Profiling
• Compound inhibits an enzymes activity
• Various concentrations compared controls used to
generate IC50s
• Substrate Selectivity Profiling
• Measure a kinases ability to phosphorylate substrates
• ~400 wild-type and ~200 mutant targets available in
human kinome
• Customized project design and data analysis with two-
week turnaround time
• Assay Offerings: ADP-Glo™ Assay, LanthaScreen™

9. ASK ME MORE
9
plobo@signalchem.com