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High throughput testing of drug library substances and monoclonal antibodies for capacity to reduce formation of cystatin C dimers to identify candidates for treatment of hereditary cystatin C amyloid angiopathy.

Östner, Gustav LU ; Lindström, Veronica LU ; Postnikov, Alexander B; Solovyeva, Tatiana I; Emilsson, Ossur I and Grubb, Anders LU (2011) In Scandinavian Journal of Clinical and Laboratory Investigation 71(8). p.676-682
Abstract
Objective. To establish a high-throughput system for testing the ability of drugs or monoclonal antibodies to reduce the in vitro formation of cystatin C dimers to identify substances potentially useful for treatment of patients with hereditary cystatin C amyloid angiopathy (HCCAA). Methods. Various combinations of incubation temperature, time period, guanidinium chloride concentration and concentration of cystatin C monomers were tested in low-volume formats to induce dimer formation of recombinant cystatin C. The extent of dimerization was analysed by gel filtration chromatography and agarose gel electrophoresis. Results. A high-throughput system based upon agarose gel electrophoresis was developed and used to test 1040 drugs in a... (More)
Objective. To establish a high-throughput system for testing the ability of drugs or monoclonal antibodies to reduce the in vitro formation of cystatin C dimers to identify substances potentially useful for treatment of patients with hereditary cystatin C amyloid angiopathy (HCCAA). Methods. Various combinations of incubation temperature, time period, guanidinium chloride concentration and concentration of cystatin C monomers were tested in low-volume formats to induce dimer formation of recombinant cystatin C. The extent of dimerization was analysed by gel filtration chromatography and agarose gel electrophoresis. Results. A high-throughput system based upon agarose gel electrophoresis was developed and used to test 1040 drugs in a clinical drug library for their capacity to reduce cystatin C dimer formation in vitro. Seventeen substances reducing dimer formation by more than 30% were identified. A similar system for testing the capacity of monoclonal antibodies against cystatin C to reduce the in vitro formation of cystatin C dimers was also developed and used to test a panel of 12 monoclonal antibodies. Seven antibodies reducing dimer formation by more than 30% were identified and the two most potent, Cyst28 and HCC3, reduced dimerization by 75 and 60%, respectively. Conclusion. We constructed a simple high-throughput system for testing the capacity of drugs and monoclonal antibodies to reduce the in vitro formation of cystatin C dimers and several candidates for treatment of HCCAA could be identified. (Less)
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organization
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type
Contribution to journal
publication status
published
subject
in
Scandinavian Journal of Clinical and Laboratory Investigation
volume
71
issue
8
pages
676 - 682
publisher
Informa Healthcare
external identifiers
  • wos:000296980600010
  • pmid:22017167
  • scopus:81255143205
ISSN
1502-7686
DOI
10.3109/00365513.2011.621026
language
English
LU publication?
yes
id
2db7ad00-f988-445a-b520-dd3283a0e2b1 (old id 2200320)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/22017167?dopt=Abstract
date added to LUP
2011-11-02 12:19:58
date last changed
2017-01-01 05:52:22
@article{2db7ad00-f988-445a-b520-dd3283a0e2b1,
  abstract     = {Objective. To establish a high-throughput system for testing the ability of drugs or monoclonal antibodies to reduce the in vitro formation of cystatin C dimers to identify substances potentially useful for treatment of patients with hereditary cystatin C amyloid angiopathy (HCCAA). Methods. Various combinations of incubation temperature, time period, guanidinium chloride concentration and concentration of cystatin C monomers were tested in low-volume formats to induce dimer formation of recombinant cystatin C. The extent of dimerization was analysed by gel filtration chromatography and agarose gel electrophoresis. Results. A high-throughput system based upon agarose gel electrophoresis was developed and used to test 1040 drugs in a clinical drug library for their capacity to reduce cystatin C dimer formation in vitro. Seventeen substances reducing dimer formation by more than 30% were identified. A similar system for testing the capacity of monoclonal antibodies against cystatin C to reduce the in vitro formation of cystatin C dimers was also developed and used to test a panel of 12 monoclonal antibodies. Seven antibodies reducing dimer formation by more than 30% were identified and the two most potent, Cyst28 and HCC3, reduced dimerization by 75 and 60%, respectively. Conclusion. We constructed a simple high-throughput system for testing the capacity of drugs and monoclonal antibodies to reduce the in vitro formation of cystatin C dimers and several candidates for treatment of HCCAA could be identified.},
  author       = {Östner, Gustav and Lindström, Veronica and Postnikov, Alexander B and Solovyeva, Tatiana I and Emilsson, Ossur I and Grubb, Anders},
  issn         = {1502-7686},
  language     = {eng},
  number       = {8},
  pages        = {676--682},
  publisher    = {Informa Healthcare},
  series       = {Scandinavian Journal of Clinical and Laboratory Investigation},
  title        = {High throughput testing of drug library substances and monoclonal antibodies for capacity to reduce formation of cystatin C dimers to identify candidates for treatment of hereditary cystatin C amyloid angiopathy.},
  url          = {http://dx.doi.org/10.3109/00365513.2011.621026},
  volume       = {71},
  year         = {2011},
}