A diabody that dissociates to monomer forms at low concentration: Effects on binding activity and tumor targeting

Bao Cheng Huang, Linda J. Foote, Trish K. Lankford, Sandra M. Davern, Cathy K. McKeown, Stephen Kennel

Research output: Contribution to journalArticle

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Abstract

A human scFv, 15-9, was selected from a phage display library for binding to murine laminin-1. A diabody was made from the scFv by shortening the linker from 15 to 5 amino acids between the VH and VL sequence. Radioiodinated scFv and diabody were analyzed for size, binding to laminin, and biodistribution in tumor bearing mice. Diabody preparations at concentrations greater than 10 nM were largely dimer forms (∼60 kDa) as judged by gel filtration, but diluted diabody was eluted as a monomer (∼30 kDa). At low concentrations the radiolabeled diabody did not bind well to laminin. The 125I diabody had significantly lower accumulation in tumors than did the scFv when injected at lower concentrations. These data indicate that the diabody dimer dissociates at concentrations of about 10 nM resulting in monomers with no binding activity for laminin and poor tumor homing properties.

Original languageEnglish (US)
Pages (from-to)999-1005
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume327
Issue number4
DOIs
StatePublished - Feb 25 2005
Externally publishedYes

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Laminin
Tumors
Monomers
Dimers
Bearings (structural)
Neoplasms
Bacteriophages
Libraries
Gel Chromatography
Gels
Display devices
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

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A diabody that dissociates to monomer forms at low concentration : Effects on binding activity and tumor targeting. / Huang, Bao Cheng; Foote, Linda J.; Lankford, Trish K.; Davern, Sandra M.; McKeown, Cathy K.; Kennel, Stephen.

In: Biochemical and Biophysical Research Communications, Vol. 327, No. 4, 25.02.2005, p. 999-1005.

Research output: Contribution to journalArticle

Huang, Bao Cheng ; Foote, Linda J. ; Lankford, Trish K. ; Davern, Sandra M. ; McKeown, Cathy K. ; Kennel, Stephen. / A diabody that dissociates to monomer forms at low concentration : Effects on binding activity and tumor targeting. In: Biochemical and Biophysical Research Communications. 2005 ; Vol. 327, No. 4. pp. 999-1005.
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