Generation and characterization of a diabody targeting the avB6 integrin
Generation and characterization of a diabody targeting the avB6 integrin
The ?v?6 integrin is up-regulated in cancer and wound healing but it is not generally expressed in healthy adult tissue. There is increasing evidence that it has a role in cancer progression and will be a useful target for antibody-directed cancer therapies. We report a novel recombinant diabody antibody fragment that targets specifically ?v?6 and blocks its function. The diabody was engineered with a C-terminal hexahistidine tag (His tag), expressed in Pichia pastoris and purified by IMAC. Surface plasmon resonance (SPR) analysis of the purified diabody showed affinity in the nanomolar range. Pre-treatment of ?v?6-expressing cells with the diabody resulted in a reduction of cell migration and adhesion to LAP, demonstrating biological function-blocking activity. After radio-labeling, using the His-tag for site-specific attachment of (99m)Tc, the diabody retained affinity and targeted specifically to ?v?6-expressing tumors in mice bearing isogenic ?v?6 +/- xenografts. Furthermore, the diabody was specifically internalized into ?v?6-expressing cells, indicating warhead targeting potential. Our results indicate that the new ?v?6 diabody has a range of potential applications in imaging, function blocking or targeted delivery/internalization of therapeutic agents.
1-9
Kogelberg, Heide
f1a1fcd6-c4bf-4355-8678-13c9256cd475
Miranda, Enrique
ff518362-5c5e-4637-b784-6bb32dade54e
Burnet, Jerome
59803900-cd01-4644-9406-a7bf44ef1fc0
Ellison, David
3ca39d61-b268-4a9b-98db-cc6f92d70e4f
Tolner, Berend
beea1fdc-6576-4000-b810-2d179e78cfcb
Foster, Julie
dccf23d1-c78e-4b66-b58c-1d53e5447aa9
Picón, Carmen
c3f20cd6-0651-4a0d-9297-74208a782c4b
Thomas, Gareth J.
2ff54aa9-a766-416b-91ee-cf1c5be74106
Meyer, Tim
28c8fc8e-873f-4278-9443-cb7c9f9f487f
Marshall, John F
bc569c17-f2b8-4085-a6ff-d4a42da43c27
Mather, Stephen J
f001f3e5-e9a2-4525-92b0-8b8d3c45d026
Chester, Kerry
c7014f1a-71d9-4ca0-959d-2164f2b9b3e2
4 September 2013
Kogelberg, Heide
f1a1fcd6-c4bf-4355-8678-13c9256cd475
Miranda, Enrique
ff518362-5c5e-4637-b784-6bb32dade54e
Burnet, Jerome
59803900-cd01-4644-9406-a7bf44ef1fc0
Ellison, David
3ca39d61-b268-4a9b-98db-cc6f92d70e4f
Tolner, Berend
beea1fdc-6576-4000-b810-2d179e78cfcb
Foster, Julie
dccf23d1-c78e-4b66-b58c-1d53e5447aa9
Picón, Carmen
c3f20cd6-0651-4a0d-9297-74208a782c4b
Thomas, Gareth J.
2ff54aa9-a766-416b-91ee-cf1c5be74106
Meyer, Tim
28c8fc8e-873f-4278-9443-cb7c9f9f487f
Marshall, John F
bc569c17-f2b8-4085-a6ff-d4a42da43c27
Mather, Stephen J
f001f3e5-e9a2-4525-92b0-8b8d3c45d026
Chester, Kerry
c7014f1a-71d9-4ca0-959d-2164f2b9b3e2
Kogelberg, Heide, Miranda, Enrique, Burnet, Jerome, Ellison, David, Tolner, Berend, Foster, Julie, Picón, Carmen, Thomas, Gareth J., Meyer, Tim, Marshall, John F, Mather, Stephen J and Chester, Kerry
(2013)
Generation and characterization of a diabody targeting the avB6 integrin.
PLoS ONE, 8 (9), .
(doi:10.1371/journal.pone.0073260).
(PMID:24023846)
Abstract
The ?v?6 integrin is up-regulated in cancer and wound healing but it is not generally expressed in healthy adult tissue. There is increasing evidence that it has a role in cancer progression and will be a useful target for antibody-directed cancer therapies. We report a novel recombinant diabody antibody fragment that targets specifically ?v?6 and blocks its function. The diabody was engineered with a C-terminal hexahistidine tag (His tag), expressed in Pichia pastoris and purified by IMAC. Surface plasmon resonance (SPR) analysis of the purified diabody showed affinity in the nanomolar range. Pre-treatment of ?v?6-expressing cells with the diabody resulted in a reduction of cell migration and adhesion to LAP, demonstrating biological function-blocking activity. After radio-labeling, using the His-tag for site-specific attachment of (99m)Tc, the diabody retained affinity and targeted specifically to ?v?6-expressing tumors in mice bearing isogenic ?v?6 +/- xenografts. Furthermore, the diabody was specifically internalized into ?v?6-expressing cells, indicating warhead targeting potential. Our results indicate that the new ?v?6 diabody has a range of potential applications in imaging, function blocking or targeted delivery/internalization of therapeutic agents.
Text
2013 Generate and characterisation of a diabody targeting the integrin.pdf
- Version of Record
More information
Accepted/In Press date: 19 July 2013
Published date: 4 September 2013
Organisations:
Cancer Sciences
Identifiers
Local EPrints ID: 395573
URI: http://eprints.soton.ac.uk/id/eprint/395573
ISSN: 1932-6203
PURE UUID: 46a3fc51-d31e-4bcd-997c-1983c88ee592
Catalogue record
Date deposited: 01 Jun 2016 13:47
Last modified: 15 Mar 2024 00:43
Export record
Altmetrics
Contributors
Author:
Heide Kogelberg
Author:
Enrique Miranda
Author:
Jerome Burnet
Author:
David Ellison
Author:
Berend Tolner
Author:
Julie Foster
Author:
Carmen Picón
Author:
Tim Meyer
Author:
John F Marshall
Author:
Stephen J Mather
Author:
Kerry Chester
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics