Preparation of Molecularly Imprinted Poly(N-Isopropylacrylamide) Thermosensitive Based Cryogels

Methods Mol Biol. 2022:2466:249-260. doi: 10.1007/978-1-0716-2176-9_17.

Abstract

Cryogels are defined as polymeric gel matrices with interconnected macropores providing an advantage to be efficient carriers enabling unhindered diffusion of interested molecule. Cryogels could be easily prepared with the combination of molecular imprinting. Molecular imprinting technology provides selective and sensitive recognition for biomolecules. Immunoglobulin G (IgG) is a main effector component in human response. It is recently well recognized that the purification strategies for IgG have intensively gained attention to treat immune defects. Several methods including affinity chromatography have been applied for the purification of IgG from complex media. The purification of IgG plays a crucial role in medical applications using several materials. Among these, cryogels have been widely applied for the purification of several biomolecules. They offer to create low-cost affinity systems with high chemical and physical stability. Above all, temperature sensitive polymers enable a reversible phase transition against small temperature changes, by the way, reversible swelling and shrinking manner is observed.In this chapter, immunoglobulin G imprinted thermosensitive poly(N-isopropylacrylamide-N methacryloyl-(l)-histidine) [p(NIPA-MAH)/IgG-MIP] monolithic cryogel is explained. The preparation and characterization of cryogels are summarized. In addition, IgG binding studies with different parameters are briefly described. Herein, an effective design principle is presented to create imprinted temperature-sensitive cryogels for IgG purification. A p(NIPA-MAH)/IgG-MIP monolithic cryogel was synthesized for IgG purification. Afterward, IgG binding capacity of p(NIPA-MAH)/IgG-MIP cryogels was examined in different experimental conditions. Apart from these, selectivity of the p(NIPA-MAH)/IgG-MIP cryogel was shown by comparing IgG binding capacity of nonimprinted [p(NIPA-MAH)/NIP] one. Finally, the IgG purification ability of the p(NIPA-MAH)/IgG-MIP cryogel from human plasma was demonstrated proving its application in affinity chromatography using real sample.

Keywords: IgG purification; Molecular imprinting; N-Isopropylacrylamide; Thermosensitive cryogel.

MeSH terms

  • Acrylamides
  • Acrylic Resins
  • Adsorption
  • Cryogels* / chemistry
  • Humans
  • Immunoglobulin G / chemistry
  • Molecular Imprinting*
  • Polymers

Substances

  • Acrylamides
  • Acrylic Resins
  • Cryogels
  • Immunoglobulin G
  • Polymers
  • poly-N-isopropylacrylamide
  • N-isopropylacrylamide