Article A Comparative Study of “Grafting to” and “Grafting from” Conjugation Methods for the Preparation of Antibody-Temperature-Responsive Polymer Conjugates

Erika Yoshihara (National Institute for Materials Science) ; Ahmed Nabil ; Michihiro Iijima ; Mitsuhiro Ebara SAMURAI ORCID (National Institute for Materials Science)

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Erika Yoshihara, Ahmed Nabil, Michihiro Iijima, Mitsuhiro Ebara. A Comparative Study of “Grafting to” and “Grafting from” Conjugation Methods for the Preparation of Antibody-Temperature-Responsive Polymer Conjugates. ACS Omega. 2024, 9 (20), 22043-22050.
SAMURAI

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(abstract)

This paper evaluates the efficiency of introducing poly (N-isopropyl acrylamide) (PNIPAAm) into immunoglobulin G (IgG) using two different conjugation methods: grafting to (GT) and grafting from (GF). The IgG-PNIPAAm conjugates were characterized using SDS-PAGE, dynamic light scattering (DLS), and temperature-responsive phase transition behavior. The SDS-PAGE analysis revealed that the GF method was more efficient in introducing the polymer than the GT method, with a calculated polymer introduction ratio of 61% and 34%, respectively. The GF method proved to be less susceptible to steric hindrance and more efficient in introducing high molecular weight polymers into proteins. Furthermore, the lower critical solubility temperature (LCST) of IgG-PNIPAAm produced by the GF method closely matches that of IgG-PNIAAm produced by a polymer with a chain length in the tens of thousands introduced by the GT method. This suggests that the molecular weight of the polymer in the conjugate produced by the GF method is also in the tens of thousands. These results are consistent with previous studies comparing the GT and GF methods on similar systems. This study represents an important step towards understanding how the choice of polymer incorporation method affects the properties of IgG-PNIPAAm conjugates. For application in simple diagnostics, we also propose using this system as an alternative method for antigen purification and enrichment.

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Keyword: Polymer-protein conjugates, Poly(N-Isopropylacrylamide), Grafting from, Antibody, Grafting to.

Date published: 2024-05-21

Publisher: American Chemical Society (ACS)

Journal:

  • ACS Omega (ISSN: 24701343) vol. 9 issue. 20 p. 22043-22050

Funding:

  • Japan Society for the Promotion of Science JP19H04476
  • Japan Society for the Promotion of Science JP20H05877

Manuscript type: Publisher's version (Version of record)

MDR DOI:

First published URL: https://doi.org/10.1021/acsomega.4c00103

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Updated at: 2024-12-04 14:53:02 +0900

Published on MDR: 2024-12-04 14:53:02 +0900

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