Glycoengineered Antibody-Drug Conjugates

The enzymes of Endo-S, PNGase-F, and AlfC were expressed in E. coli following the reported procedures^{8 (link), 37 (link), 42 (link), 43 (link)}. The glycoengineered azido-herceptin (7) and DBCO-MMAE (8) were prepared following our previous paper^{8 (link)}. DM1 (N₂′-deacetyl-N₂′-(3-mercapto-1-oxopropyl)- maytansine), MMAE (monomethyl auristain E), and SMCC (N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate) were purchased from Levena Biopharma (Nanjing, China). Other chemical reagents and solvents were purchased from Sinopharm Chemical Reagent Co. (Shanghai, China) or Sigma-Aldrich (Shanghai, China) and used without further purification. The MAbPac RP column (4 μm, 3.0 × 100mm) was purchased from ThermoFisher. Nuclear magnetic resonance (NMR) spectra were measured on a Varian-MERCURY Plus-400 or 500 instrument. ESI-HRMS spectra were measured on an Agilent 6230 LC-TOF MS spectrometer.

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Tang Y., Tang F., Yang Y., Zhao L., Zhou H., Dong J, & Huang W. (2017). Real-Time Analysis on Drug-Antibody Ratio of Antibody-Drug Conjugates for Synthesis, Process Optimization, and Quality Control. Scientific Reports, 7, 7763.

Publication 2017

MAbPac RP column MERCURY Plus-400 Agilent 6230 LC-TOF MS spectrometer

Cyclohexane E coli Endo Enzymes Herceptin Maytansine Mercury Nuclear magnetic resonance Pngase f Solvents

Corresponding Organization :

Other organizations : Shenyang Pharmaceutical University, Center for Excellence in Molecular Cell Science, Shanghai Institute of Materia Medica, Chinese Academy of Sciences

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Variable analysis

independent variables

Expression of the enzymes Endo-S, PNGase-F, and AlfC in E. coli
Preparation of glycoengineered azido-herceptin (7) and DBCO-MMAE (8)

dependent variables

Not explicitly mentioned

control variables

Use of reported procedures for enzyme expression
Use of previous paper for preparation of glycoengineered compounds
Purchasing of reagents and solvents from specified suppliers without further purification
Use of specific analytical equipment (MAbPac RP column, Varian-MERCURY Plus-400 or 500 NMR instrument, Agilent 6230 LC-TOF MS spectrometer)

controls

Positive control: Not specified
Negative control: Not specified

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