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 J. Kennel

doi:10.1016/j.bbrc.2004.12.114

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 J. Kennel

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

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 language	English
Pages (from-to)	999-1005
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	327
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2004.12.114
State	Published - Feb 25 2005
Externally published	Yes

Funding

We thank Drs. Peter Hoyt and Dabney Johnson who helped in editing the manuscript. This work was supported by funding from the Medical Sciences Division of US DOE contract AC005-00OR22725 under Grant ERKP038. The generous contribution of the phage libraries from The MRC Laboratory of Molecular Biology and the MRC Centre for Protein Engineering (Cambridge, UK) is gratefully acknowledged.

Funders	Funder number
Medical Sciences Division of US DOE	ERKP038, AC005-00OR22725

Keywords

Diabody
Dissociation
Tumor targeting

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.bbrc.2004.12.114

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

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.",

keywords = "Diabody, Dissociation, Tumor targeting",

author = "Huang, {Bao Cheng} and Foote, {Linda J.} and Lankford, {Trish K.} and Davern, {Sandra M.} and McKeown, {Cathy K.} and Kennel, {Stephen J.}",

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T2 - Effects on binding activity and tumor targeting

AU - Huang, Bao Cheng

AU - Foote, Linda J.

AU - Lankford, Trish K.

AU - Davern, Sandra M.

AU - McKeown, Cathy K.

AU - Kennel, Stephen J.

PY - 2005/2/25

Y1 - 2005/2/25

N2 - 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.

AB - 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.

KW - Diabody

KW - Dissociation

KW - Tumor targeting

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 4

ER -

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

Abstract

Funding

Keywords

UN SDGs

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