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.

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.

• Authors: Gustav Östner, Veronica Lindström, Alexander B Postnikov, Tatiana I Solovyeva, Össur I Emilsson, Anders Grubb
• Journal: Scandinavian journal of clinical and laboratory investigation (Scand J Clin Lab Invest) Vol. 71 Issue 8 Pg. 676-82 (Dec 2011) ISSN: 1502-7686 [Electronic] England
• PMID: 22017167 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Cystatin C
• Recombinant Proteins
• Small Molecule Libraries
• Solutions
• Guanidine

Topics

• Antibodies, Monoclonal (chemistry, metabolism, pharmacology)
• Cerebral Amyloid Angiopathy (congenital, drug therapy, metabolism, physiopathology)
• Cerebral Arteries (drug effects, metabolism, physiopathology)
• Chromatography, Gel
• Cystatin C (antagonists & inhibitors, metabolism)
• Dimerization
• Drug Discovery (methods)
• Electrophoresis, Agar Gel
• Guanidine (adverse effects)
• High-Throughput Screening Assays
• Humans
• Recombinant Proteins (antagonists & inhibitors, metabolism)
• Small Molecule Libraries (chemistry, metabolism, pharmacology)
• Solutions