Title

Unexpected side products in the conjugation of an amine-derivatized morpholino oligomer with p-isothiocyanate benzyl DTPA and their removal

UMMS Affiliation

Department of Radiology, Division of Nuclear Medicine; Department of Medicine

Date

2-1-2011

Document Type

Article

Medical Subject Headings

Amines; Animals; Base Sequence; Chemical Fractionation; DNA; Isothiocyanates; Isotope Labeling; Mice; Morpholines; Pentetic Acid; Phosphorus Compounds; Polymers

Disciplines

Radiochemistry | Radiology

Abstract

In connection with pretargeting, an amine-derivatized morpholino phosphorodiamidate oligomer (NH(2)-cMORF) was conjugated conventionally with p-isothiocyanate benzyl-DTPA (p-SCN-Bn-DTPA). However, after (111)In radiolabeling, unexpected label instability was observed. To understand this instability, the NH(2)-cMORF and, as control, the native cMORF without the amine were conjugated in the conventional manner. Surprisingly, the (111)In labeling of the native cMORF conjugate was equally effective as that of the NH(2)-cMORF conjugate (>95%) despite the absence of the amine group. Furthermore, heating the radiolabeled NH(2)-cMORF and native cMORF conjugates resulted in a 35% loss and a complete loss of the label, respectively. Since the (111)In labeled DTPA is known to be stable, the instability in both cases must be due to some unstable association of DTPA to the cMORF, presumably unstable association to some endogenous sites in cMORF. Based on this assumption, a postconjugation-prepurification heating step was introduced, and labeling efficiency and stability were again investigated. By introducing the heating step, the side products were dissociated, and after purification and labeling, the NH(2)-cMORF conjugate provided a stable label and high labeling efficiency with no need for postlabeling purification. The biodistribution of this radiolabeled conjugate in normal mice showed significantly lower backgrounds compared with the labeled unstable native cMORF conjugate. In conclusion, the conventional conjugation procedure to attach the p-SCN-Bn-DTPA to NH(2)-cMORF resulted in side product(s) that were responsible for the (111)In label instability. Adding a postconjugation-prepurification heating step dissociated the side products, improved the label stability and lowered tissue backgrounds in mice.

Rights and Permissions

Citation: Nucl Med Biol. 2011 Feb;38(2):159-63. doi: 10.1016/j.nucmedbio.2010.08.008. Link to article on publisher's site

Related Resources

Link to Article in PubMed

Keywords

Chelator, Conjugation, Radiolabeling, DNA analogues

PubMed ID

21315270