Conjugation Guidelines

Conjugation Chemistry Determination Flow Diagram (95 K PDF)

There are 3 items to consider when determining peptide conjugation

1. Carrier Protein: attachment of the peptide segment to a carrier protein is a key factor in eliciting an immune response.
   1. KLH: Keyhole limpet hemocyanin is derived from marine mollusk
   2. BSA: Bovine serum albumin is derived from cows
   3. RSA: Rabbit serum albumin is derived from rabbits
   4. OVA: Ovalbumin is derived from egg whites
   
   
2. Location of peptide segment within the native protein: necessary in determining the appropriate conjugation chemistry of the peptide to the carrier protein for presenting the peptide segment to the immune system.
   1. N-terminal region of the protein (within the first 100 amino acids)
   2. Internal region (between the N-terminal and C-terminal regions)
      
   3. C-terminal region of the protein (within the last 100 amino acids)
   
   
3. Conjugation Chemistry: the attachment site of the peptide to the carrier protein
   1. EDC: peptide attachment to carrier protein via the carboxyl groups within the peptide sequence (D, E, and C-terminal carboxyl group)
   2. Activated EDC: peptide attachment to carrier protein via the amino groups within the peptide sequence (K and N-terminal amino group)
   3. MBS: peptide attachment to carrier protein via the thiol group of a cysteine residue within the peptide sequence.