Citation: Xia Liu, Wenzhuo Dong, Mengqian Jia, Dexiu Zhang, Jingyi Niu, Jiwei Shen, Chaozhan Wang, Yinmao Wei. Two-site synergistic binding strategy for improving adsorption and separation performance of antibodies[J]. Chinese Chemical Letters, ;2025, 36(12): 110991. doi: 10.1016/j.cclet.2025.110991 shu

Two-site synergistic binding strategy for improving adsorption and separation performance of antibodies

Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, College of Chemistry & Materials Science, Northwest University, Xi'an 710127, China

jiweish@nwu.edu.cn

ymwei@nwu.edu.cn

Received Date: 13 September 2024
Revised Date: 27 December 2024
Accepted Date: 20 February 2025
Available Online: 20 February 2025

Figures(5)

The adsorption and separation of antibody drugs are of great significance, but the promising hydrophobic charge induction chromatography (HCIC) and boronate affinity chromatography (BAC) suffer from low specific due to the limitations of single-site adsorption mechanism as well as low adsorption capacity of adsorbents, resulting in a lower purity and recovery of antibodies. To address this issue, this work proposes a two-site synergistic binding strategy integrating HCIC and BAC mechanism on a polymer brushes-grafted adsorbent. Five adsorbents were easily created by polymerizing the mixed monomers of 5-acryloylaminobenzimidazole, 3-acryloylamide phenylboronic acid and acrylamide on surface of agarose gel via activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP). The molecular docking implies that the two-site synergistic binding towards immunoglobulin G (IgG) originates from the closely adjacent boronic and benzimidazole side groups in the polymer chains with monomer ratio of 1:1:0. The inference was verified by the effect of three monomer ratios and adsorption conditions on the adsorption performance of IgG. The adsorbent with two-site synergy possesses an excellent specific, enhanced affinity (K_d = 3.9 × 10⁻⁶ mol/L) and adsorption capacity (Q_m = 253 mg/g) towards IgG. Benefiting from the advantages, IgG from serum and monoclonal antibody (mAb) from cell culture achieve purities of 95.8% and 98.3%, and recoveries of 95.7% and 97.5%, respectively. The results are comparable to those with protein A adsorbent considered to have the best specific so far, indicating the potential of the two-site synergistic binding strategy in the purification of antibody drugs.
- Boronate affinity chromatography,
- Hydrophobic charge induction chromatography,
- Polymer brushes-grafted resin,
- Two-site synergistic adsorption,
- Antibody
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