1. Programme analysis
Timeline
Deliverables
Optimization of codon
Fusion of tag
Purification methods
Optional host cells
2. Expression System System Selection (Please skip this step if you can provide an expression strain.)
Optional expression vector
pFastBac1 of secretory and intracellular types, with a label at the N- or C-terminus.
Optional expression strain
Optional expression host Spodoptera frugiperda (Spodoptera frugiperda) ovarian cells, SF9 SF21 High5
3. Vector construction
Timeline
Deliverables
A. Gene optimization and synthesis
B. Subcloned into pFasbac1
C. Preparation of recombinant Bacmid
Synthesis & subcloning: 2-3 weeks
Bacmid Preparation: 1-2 weeks
1. Enzyme digestion/sequencing report
2. DH10 containing expression plasmid
3. Expression plasmids
4. Pellet and P1 Generation Virus Preparation
Timeline
Deliverables
D. P1 generation Baculovirus preparation
E. P1 transfection WB identification
P1 generation and identification: 1-2 weeks
5. Pellet and P2 Generation Virus Preparation
Timeline
Deliverables
F. P2 generation Baculovirus preparation
G. P2 infection WB identification
P1 generation and identification: 1-2 weeks
4. Certification report of P2 generation
6. Sample purification
Timeline
Deliverables
H. Ni column affinity purification
I. Purity detection
Expression & purification: 1-2 weeks
5. Protein sample
(50-500 ug protein available)
7. Scale-up culture and protein purification
Timeline
Deliverables
J. Scale up (according to customer demand)
K. Ni column affinity purification
L. Purified protein detection
M. Protein shipment
Amplification culture: 2-3 weeks
Protein purification: 1-2 weeks
6. Protein purity identification report
7. Finished protein
(According to the expression of small test results agreement)
Supplementary explanation
We use insect cell systems to obtain proteins. In general, signal peptides will be added to allow the mature protein to be secreted into the culture medium. However, not all proteins can be secreted or expressed as expected. On the basis of respecting scientific facts, we cannot promise that all proteins can be successfully expressed or expressed in a secreted form. We also do not promise that the protein obtained will have the expected physiological activity of the customer.