Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria): La. gel antibodies against proteins (transferrins)

Anikó Takátsy, Anikó Kilár, Ferenc Kilár, Stellan Hjertén

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Artificial antibodies in the form of gel granules were prepared by the molecular imprinting technique from the monomers acrylamide and N,N′-methylene-bis-acrylamide. Gel granules, freed from the selectively adsorbed protein (the antigen), are neutral and, accordingly, do not migrate in an electrical field. However, upon selective interaction with the antigen at a pH different from its pl, the granules become charged. The selectivity of the gel antibodies was studied by free zone electrophoresis in a tube with inside diameter larger than the size of the granules. Such electrophoretic analyses showed that gel antibodies against iron-free transferrin had a high selectivity for this protein, although some crossreaction took place with iron-saturated transferrin, indicating that these artificial antibodies can easily distinguish the minute differences in the 3-D structure of the transferrins. Analogously, gel antibodies against iron-saturated transferrin were highly selective for this protein with some crossreaction with iron-free transferrin. The mobilities of iron-free and iron-saturated transferrin are very similar, and, therefore, capillary free zone electrophoresis cannot distinguish between these structurally related proteins. However, significant differences in the mobilities of the selective gel granules can be observed depending on their interaction with iron-free or iron-saturated transferrin, i.e., the artificial gel antibodies may become powerful analytical tools.

Original languageEnglish
Pages (from-to)2802-2809
Number of pages8
JournalJournal of separation science
Volume29
Issue number18
DOIs
Publication statusPublished - Dec 1 2006

Keywords

  • Artificial antibodies
  • Free zone electrophoresis
  • Molecular imprinting
  • Molecular recognition
  • Transferrin

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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