How does CPC conjugate my peptide to a carrier protein for antibody production?
When producing peptide-antibodies, it is important to use a carrier protein to elicit a better response from the animal. Commonly used carrier proteins are BSA and KLH, with KLH preferred for its higher immunogenicity. Peptides coupled with BSA will generate antibodies against both the peptide and the BSA, which could result in incorrect ELISA readings. For coupling carrier proteins to peptides, CPC recommends incorporating an N terminus cysteine for this reaction due to the high efficiency and proven effectiveness. For coupling through cysteine, we use MBS (maleimidobenzoic acid-N-hydroxysuccinimide ester) as a crosslinker. The initial step is to activate the protein with MBS, and then remove the excess crosslinker by SEC. This activated carrier protein is then coupled to the cysteine in the peptide through a disulfide bond with an efficiency of >95%. The only impurity in this reaction is salt formation, which CPC removes through gel filtration. As an alternative to coupling through c
When producing peptide-antibodies, it is important to use a carrier protein to elicit a better response from the animal. Commonly used carrier proteins are BSA and KLH, with KLH preferred for its higher immunogenicity. Peptides coupled with BSA will generate antibodies against both the peptide and the BSA, which could result in incorrect ELISA readings. For coupling carrier proteins to peptides, CPC recommends incorporating an N terminus cysteine for this reaction due to the high efficiency and proven effectiveness. For coupling through cysteine, we use MBS (maleimidobenzoic acid-N-hydroxysuccinimide ester) as a crosslinker. The initial step is to activate the protein with MBS, and then remove the excess crosslinker by SEC. This activated carrier protein is then coupled to the cysteine in the peptide through a disulfide bond with an efficiency of >95%. The only impurity in this reaction is salt formation, which CPC removes through gel filtration.
