New synthesis of immunogenic N6-isopent-2-enyladenosine-protein conjugate. Preparation, purification, and specificity of related antibodies

Bioconjug Chem. 1992 Jan-Feb;3(1):14-9. doi: 10.1021/bc00013a002.

Abstract

An original procedure which preserves the structure of the sugar ring is described to link a plant hormone as N6-isopent-2-enyladenosine [( 9R]iP) onto a protein carrier to prepare a more specific immunogen. This cytokinin is bound to bovine serum albumin (BSA) and ovalbumin by a five-step procedure. These [9R]iP-protein conjugates have a maximal absorption at 269 nm and show molar ratios of hormone bound to proteins in the range of 12:1 and 18:1 for BSA and ovalbumin, respectively. Polyclonal antibodies were raised in rabbits against [9R]iP-BSA and were purified by affinity chromatography. Titers and specificity of the antisera and purified antibodies were determined by ELISA and RIA. These antibodies are highly specific for [9R]iP and do not cross-react with zeatin and ribosylzeatin. An immunoaffinity matrix was prepared with a capacity of 1 microgram of [9R]iP/mL of gel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Antibodies / isolation & purification
  • Antibody Specificity
  • Chromatography, Affinity
  • Cross Reactions
  • Cytokinins / chemistry
  • Cytokinins / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Isopentenyladenosine / chemistry
  • Isopentenyladenosine / immunology*
  • Ovalbumin / chemistry
  • Ovalbumin / immunology*
  • Rabbits
  • Serum Albumin, Bovine / chemistry
  • Serum Albumin, Bovine / immunology*
  • Spectrophotometry, Ultraviolet

Substances

  • Antibodies
  • Cytokinins
  • Serum Albumin, Bovine
  • Isopentenyladenosine
  • Ovalbumin