A novel 111In-labeled antisense DNA probe with multi-chelating sites (MCS-probe) showing high specific radioactivity and labeling efficiency

Nucl Med Biol. 1999 Jan;26(1):17-21. doi: 10.1016/s0969-8051(98)00058-4.

Abstract

A multi-aminolinked oligodeoxynucleotide (ODN) was synthesized by substitution of dT with aminolinked dU in the sequence, following conjugation with isothiocyanobenzyl-EDTA (IBE) for 111In labeling. As a model target gene, the c-erbB-2 protooncogene was used. The probability of the number of aminolinked dU in the 20mer ODN was 5, but there were actually 3 and 4 in the selected antisense and sense ODNs, respectively. The IBE/ODN conjugation levels of probes with multi-chelating sites (MCS-probe) were 1.6 (antisense) and 2.4 (sense), more than 50 times higher than those of our previous studies using 5'-end aminolinked ODNs (IBE/ODN = 0.03). Labeling studies using the MCS-probe and 111In indicated that specific radioactivity as high as 48 MBq/nmol could be obtained with a labeling efficiency of over 90%. The 111In-antisense-MCS-probe could bound to sense ODN under physiological conditions, but the 111In-sense-MCA-probe could not. Thus, side-chain modification of ODN for metal labeling is considered to be useful for antisense techniques.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antisense Elements (Genetics)
  • DNA, Antisense / chemical synthesis*
  • DNA, Antisense / metabolism
  • Edetic Acid / analogs & derivatives
  • Indium Radioisotopes*
  • Isothiocyanates
  • Isotope Labeling
  • Oncogene Proteins
  • RNA, Messenger

Substances

  • Antisense Elements (Genetics)
  • DNA, Antisense
  • Indium Radioisotopes
  • Isothiocyanates
  • Oncogene Proteins
  • RNA, Messenger
  • isothiocyanobenzyl ethylenediamine tetracetic acid
  • Edetic Acid