Affibody Molecule Conjugation Protocol

The amino acid sequence of affibody molecule Z_hcHER2:342 was MIHHHHHHLQVDNFNKEMRNAYWEIALLPNLNNQQKRAIRSLYDDSQSANLLAEAKKLNDAQAPKVDC.
The cF₁₃-affibody conjugate was constructed according to our previous report [38 (link),39 (link)], and the synthetic route is described in Scheme A2. The obtained DNA-affibody conjugate was analyzed and characterized by UV-vis spectroscopy and 2% agarose gel electrophoresis. The molecular weights of cF₁₃-affibody and affibody were also detected by gel permeation chromatography (GPC, Waters 2695, Milford) using polystyrene as standard. The pure cF₁₃-affibody was concentrated using Amicon ultracentrifugal filters (MWCO 10 kDa, Merck Millipore, Darmstadt, Germany) and stored at 4 °C.

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Zhang C., Fu S., Zhang F., Han M., Wang X., Du J., Zhang H, & Li W. (2022). Affibody Modified G-quadruplex DNA Micelles Incorporating Polymeric 5-Fluorodeoxyuridine for Targeted Delivery of Curcumin to Enhance Synergetic Therapy of HER2 Positive Gastric Cancer. Nanomaterials, 12(4), 696.

Publication 2022

Waters 2695 Amicon ultracentrifugal filters

Agarose gel electrophoresis Amino acid sequence Gel permeation chromatography Polystyrene Spectroscopy

Corresponding Organization :

Other organizations : Ministry of Education of the People's Republic of China, Hebei University, Hengshui University

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Variable analysis

independent variables

Conjugation of cF13 to affibody molecule Z(hcHER2:342)

dependent variables

Molecular weight of cF13-affibody conjugate
Molecular weight of affibody
UV-vis spectroscopy analysis of DNA-affibody conjugate
Agarose gel electrophoresis analysis of DNA-affibody conjugate

control variables

Polystyrene standard used for gel permeation chromatography (GPC) analysis
Amicon ultracentrifugal filters (MWCO 10 kDa) used for concentrating the purified cF13-affibody

controls

No positive or negative controls were explicitly mentioned in the information provided.

Annotations

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