I want to purify Peptide-Fc Conjugate through Affinity purification, So I tried using Mab Select Sure which is a Protein A column used for purification of mAb's, But during elution there was no peak observed, So I would like to know which column should be preferred for purification of Peptide-Fc Conjugate.
Also I'd like to know what are the possible reasons for un binding of peptide-Fc conjugate to the column ?
Dear Vikram Kumar
which kind of Fc sequence did you use?
- human, murine? Which class?
What do you mean for conjugate? Fused? At N- or C-term or chemically conjugate.
Best regards
Manuele
Protein A binds to antibodies through the Fc domain, so, if your peptide binds to the Fc region which interacts with protein A, there will be no binding.
cheers
Manuele Martinelli Mine is GLP-1 peptide covalently linked to a human IgG4-Fc heavy chain by a small peptide linker.
You may target kappa and lambda chains to affinity purify. Since the Fc region is altered and if the purification protocol was appropriately applied, this is the highly specific way to get your GLP-linked heavy chain after reduction (by cleaving the disulfides)...Look for cytiva and thermo for class-specific kappa or lambda affinity matrix to supply. Otherwise, anti-GLP-1 antibody and conjugation chemistry will help...
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