Affinity Purified Two Peptide
Antibody Package
(Peptides are designed for different regions of the
same
protein)
1. Peptide design (against 2 different epitopes of
the same protein)
Consideration for -
- Antigenicity,
N terminal and C terminal options
- Solubility (hydrophilicity
& hydrophobicity)
- Structural concerns, interactions
and presence of a.acid residues
- Specificity/cross reactivity
– paralogs, orthologs
- Bioinformatics
-
Jameson & Wolf combination
surface probability(1988)
- Homology (Blast
Analysis)
Recommendation for multiple peptides
Selection of 2 peptides based on assay needs (free consultation with
customers)
2. Peptide
synthesis and preparation
- >90%
pure, 10mgs of each peptide
- Conjugation
of peptides
3. Antibody
development (Polyclonal)
- Number of
rabbits: 4 (2 for each peptide)
- Protocol -
4
injections/3
bleeds
- Affinity Purification - Large, 1 pass
-
ELISA - 2 plates
4. Deliverables:
- Project schedule
- 1.5 – 9.0 mgs purified
antibody per peptide
- Sterile filtered
- SDS page gel
- Flow Through
- Columns
- Final project report
- Peptide balance with project term
Total project turnaround - 88 business days, 123 calendar days