title: An improved protocol for coupling synthetic peptides to carrier proteins for antibody production using DMF to solubilize peptides.
abstract: We present an improved protocol for coupling synthetic peptides to carrier proteins. In this protocol, dimethyl-formamide is used as the solvent to solubilize peptides instead of phosphate-buffered saline (PBS) or 6 M guanidine-HCl/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 m-maleimidobenzoyl-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-HCl.
authors.affiliation: Protein Research Laboratory, Research Resources Center, University of illinois at Chicago, 835 S. Wolcott Ave, Chicago, IL, USA.
authors.forename: Syed Salman
authors.initials: SS
authors.lastname: Lateef
authors.forename: Shalini
authors.initials: S
authors.lastname: Gupta
authors.forename: Lasanthi P
authors.initials: LP
authors.lastname: Jayathilaka
authors.forename: Sangeeth
authors.initials: S
authors.lastname: Krishnanchettiar
authors.forename: Jin-Sheng
authors.initials: JS
authors.lastname: Huang
authors.forename: Bao-Shiang
authors.initials: BS
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D004126:Dimethylformamide; D007136:Immunoglobulins / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology / Q000378:metabolism; D012995:Solubility
chemical_list: D002352:Carrier Proteins; D007136:Immunoglobulins; D010455:Peptides; D004126:Dimethylformamide