Totaro Kyle A, Liao Xiaoli, Bhattacharya Keshab, Finneman Jari I, Sperry Justin B, Massa Mark A, Thorn Jennifer, Ho Sa V, Pentelute Bradley L
Department of Chemistry, Massachusetts Institute of Technology , 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
Pfizer Worldwide Research and Development , 700 Chesterfield Parkway West, Chesterfield, Missouri 63017, United States.
Bioconjug Chem. 2016 Apr 20;27(4):994-1004. doi: 10.1021/acs.bioconjchem.6b00043. Epub 2016 Mar 24.
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.
1-乙基-3-(3-(二甲氨基)丙基)碳二亚胺(EDC)生物共轭反应已被用于制备医学研究用变体。虽然在优化水相应用反应方面取得了进展，但在确定干扰产物形成的条件和副反应方面关注有限。我们对EDC/N-羟基磺基琥珀酰亚胺(sNHS)介导的羧化肽和小蛋白生物共轭反应进行了系统研究。我们鉴定出了试剂和底物产生的尚未报道的副产物。本研究中使用的模型肽表明，特定底物比其他底物更容易发生副反应。从我们的研究中发现，高浓度的胺亲核试剂能使生物共轭反应更有效，但sNHS并非如此。对模型亲和体蛋白进行生物共轭反应表明，模型肽确定的趋势在更复杂的系统中也成立。
