title: Systematic Investigation of EDC/sNHS-Mediated Bioconjugation Reactions for Carboxylated Peptide Substrates.
abstract: 1-Ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC) bioconjugations have been utilized in preparing variants for medical research. While there have been advances in optimizing the reaction for aqueous applications, there has been limited focus toward identifying conditions and side reactions that interfere with product formation. We present a systematic investigation of EDC/N-hydroxysulfosuccinimide (sNHS)-mediated bioconjugations on carboxylated peptides and small proteins. We identified yet-to-be-reported side products arising from both the reagents and substrates. Model peptides used in this study illustrate particular substrates are more susceptible to side reactions than others. From our studies, we found that bioconjugations are more efficient with high concentrations of amine nucleophile but not sNHS. Performing bioconjugations on a model affibody protein show that the trends established with model peptides hold for more complex systems.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Totaro
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liao
authors.affiliation: Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
authors.forename: Keshab
authors.initials: K
authors.lastname: Bhattacharya
authors.affiliation: Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
authors.forename: Jari I
authors.initials: JI
authors.lastname: Finneman
authors.affiliation: Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
authors.forename: Justin B
authors.initials: JB
authors.lastname: Sperry
authors.affiliation: Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Massa
authors.affiliation: Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Thorn
authors.affiliation: Pfizer Worldwide Research and Development , 1 Burtt Road, Andover, Massachusetts 01810, United States.
authors.forename: Sa V
authors.initials: SV
authors.lastname: Ho
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
authors.forename: Bradley L
authors.initials: BL
authors.lastname: Pentelute
mesh_terms: D002234:Carbodiimides / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D010455:Peptides / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: C518491:1-ethyl-3-(3-(diethylamino)propyl)carbodiimide; D002234:Carbodiimides; D010455:Peptides