Peptide Synthesis
In peptide-antibody production, carrier proteins are essential for eliciting a strong immune response. KLH is preferred over BSA due to its higher immunogenicity. Using BSA may generate antibodies against both the peptide and BSA, potentially skewing ELISA res
In peptide-antibody production, carrier proteins are essential for eliciting a strong immune response. KLH is preferred over BSA due to its higher immunogenicity. Using BSA may generate antibodies against both the peptide and BSA, potentially skewing ELISA results.
For effective coupling, we recommend adding an N-terminal cysteine to the peptide. This allows for site-specific conjugation using MBS (maleimidobenzoic acid-N-hydroxysuccinimide ester) as a crosslinker. The process involves activating the carrier protein with MBS, removing excess crosslinker via SEC, then coupling to the peptide’s cysteine through a disulfide bond—achieving >95% efficiency. The only byproduct, salt, is removed by gel filtration.
As an alternative, peptides can be coupled to carrier proteins via amino groups using glutaraldehyde. This method typically yields ~80% efficiency. However, careful control of crosslinker concentration is critical to avoid unwanted protein–protein or peptide–peptide crosslinking.