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Improved method for preparing N-hydroxysuccinimide ester-containing polymers for affinity chromatography

M Wilchek1, K L Knudsen, T Miron

  • 1Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, Israel.

Bioconjugate Chemistry
|September 1, 1994
PubMed
Summary
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N,N,N

Area of Science:

  • Polymer chemistry
  • Bioconjugation techniques

Background:

  • Conventional methods for carboxyl group activation and N-hydroxysuccinimide ester formation on polymers can lead to unstable gels.
  • Hydroxy-containing resins require specific activation methods for efficient functionalization.

Purpose of the Study:

  • To introduce N,N,N',N'-Tetramethyl(succinimido) uronium tetrafluoroborate as a superior reagent for polymer functionalization.
  • To demonstrate the reagent's effectiveness in forming stable N-hydroxysuccinimide esters on various polymer supports.

Main Methods:

  • Utilizing N,N,N',N'-Tetramethyl(succinimido) uronium tetrafluoroborate for carboxyl group activation on hydroxy-containing resins.
  • Coupling ligands and proteins to the activated polymer matrices.

Main Results:

Related Experiment Videos

  • The reagent effectively activates carboxyl groups and forms N-hydroxysuccinimide esters on polymers like Sepharose, cellulose, and dextran.
  • High yields were observed for both activation and subsequent ligand/protein coupling.
  • Resulting functionalized columns demonstrated suitability for affinity chromatography and protein immobilization.

Conclusions:

  • N,N,N',N'-Tetramethyl(succinimido) uronium tetrafluoroborate offers a robust and efficient method for polymer functionalization.
  • This reagent overcomes limitations of conventional methods, enabling high-capacity affinity matrices and protein immobilization.