An Improved Protocol for Coupling Synthetic Peptides to Carrier Proteins for Antibody Production Using DMF to Solubilize Peptides 

 -  Journal of BioMolecular Techniques, 18(3), p.173-176, 2007 .
<br/><br/> We present an improved protocol for coupling synthetic peptides to carrier proteins. In this protocol, dimethylformamide is used as the solvent to solubilize peptides instead of phosphate-buffered saline (PBS)or 6 M guanidine-HC l/0.01 M phosphate buffer (pH 7). Additionally, the last desalting or dialyzing step to remove uncoupled peptides as in the traditional method is eliminated. Finally, 3 mL of 0.1 M ammonium bicarbonate is added to the carrier protein conjugated peptide solution to help the lyophilization process. Coupling of Cys-containing synthetic peptides to keyhole limpet hemocyanin or bovine serum albumin using mmaleimidobenzoyl- N-hydroxysuccinimide ester are used as the test cases. This method produces high-quality antipeptide antibodies. Also, compared to the traditional method, this procedure is simpler and useful for peptides with solubility problems in PBS or 6 M guanidine-HC l. 
<br/><br/><label>Subjects--Topical Terms: </label><br/>PEPTIDE<br/>ANTIBODY<br/>KLH<br/>BSA<br/>MBS<br/>DMF<br/>MALDI- TOF MS