Inventors
Arnon Rosenthal and Michael Leviten
Assignees
- ANNEXON, INC. (South San Francisco, CA, US)
Abstract
The present invention provides anti-C1q antibodies and methods for using the same.
CovalX Technology Used
Epitope Mapping
HM3
MALDI-ToF
Complex Tracker
Outcomes
In order to determine whether or not the CIq peptides that had been generated by proteolysis competed for binding of CIq to the antibody, analysis was performed. 25 μl of the CIq antigen (10 μM) was digested with immobilized pepsin (5 μM) and incubated at room temperature for 30 minutes. The incubated sample was centrifuged and the supernatant was removed by pipetting. To control the completion of proteolysis and optimize it to the 1000 to 3500 Da range, a MALDI mass spectrometer was used in linear mode. 5 μl of antigen peptides that were generated from the proteolysis were mixed with 5 μl of ANN-001 or ANN-005 (5 μM) before being incubated for 6 hours at 37 °C. 5 μl of the incubated mixture was mixed with 5 μl of the C1Q antigen (4 μM). This mixture contained either 2 μM/2 μM/2.5 μM of C1QA antigen/4A4B11 or MI/C1Q antigen peptides. The mixture was spotted on a MALDI plate and analyzed using a mass spectrometer that had been modified with a CovalX HM3 detection system. The data was smoothed and subtracted using the CovalX Complex Tracker Software.