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.

Summary, in English

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: Gustav Östner; Veronica Lindström; Alexander B Postnikov; Tatiana I Solovyeva; Ossur I Emilsson; Anders Grubb
• Department/s: Division of Clinical Chemistry and Pharmacology
• Publishing year: 2011
• Language: English
• Pages: 676-682
• Publication/Series: Scandinavian Journal of Clinical and Laboratory Investigation
• Volume: 71
• Issue: 8
• Full text: - 173 kB; Download statistics
• Links: http://www.ncbi.nlm.nih.gov/pubmed/22017167?dopt=Abstract; http://dx.doi.org/10.3109/00365513.2011.621026
• Document type: Journal article
• Publisher: Informa Healthcare
• Topic: Pharmacology and Toxicology; Medicinal Chemistry
• Status: Published
• ISBN/ISSN/Other: ISSN: 1502-7686