Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma
Decysin-1 (ADAMDEC1) is an orphan ADAM-like metalloprotease with unknown biological function and a short domain structure. ADAMDEC1 mRNA has previously been demonstrated primarily in macrophages and mature dendritic cells. Here, we generated monoclonal antibodies (mAbs) against the mature ADAMDEC1 p...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Journal article |
| Language: | English |
| Published: |
Taylor and Francis
2017
|
| _version_ | 1826292306520571904 |
|---|---|
| author | Lund, J Elimar Bitsch, A Grønbech Rasch, M Enoksson, M Troeberg, L Nagase, H Loftager, M Overgaard, M Petersen, H |
| author_facet | Lund, J Elimar Bitsch, A Grønbech Rasch, M Enoksson, M Troeberg, L Nagase, H Loftager, M Overgaard, M Petersen, H |
| author_sort | Lund, J |
| collection | OXFORD |
| description | Decysin-1 (ADAMDEC1) is an orphan ADAM-like metalloprotease with unknown biological function and a short domain structure. ADAMDEC1 mRNA has previously been demonstrated primarily in macrophages and mature dendritic cells. Here, we generated monoclonal antibodies (mAbs) against the mature ADAMDEC1 protein, as well as mAbs specific for the ADAMDEC1 pro-form, enabling further investigations of the metalloprotease. The generated mAbs bind ADAMDEC1 with varying affinity and represent at least six different epitope bins. Binding of mAbs to one epitope bin in the C-terminal disintegrin-like domain efficiently reduces the proteolytic activity of ADAMDEC1. A unique mAb, also recognizing the disintegrin-like domain, stimulates the caseinolytic activity of ADAMDEC1 while having no significant effect on the proteolysis of carboxymethylated transferrin. Using two different mAbs binding the disintegrin-like domain, we developed a robust, quantitative sandwich ELISA and demonstrate secretion of mature ADAMDEC1 protein by primary human macrophages. Surprisingly, we also found ADAMDEC1 present in human plasma with an approximate concentration of 0.5 nM. The presence of ADAMDEC1 both in human plasma and in macrophage cell culture supernatant were biochemically validated using immunoprecipitation and Western blot analysis demonstrating that ADAMDEC1 is secreted in a mature form. |
| first_indexed | 2024-03-07T03:12:39Z |
| format | Journal article |
| id | oxford-uuid:b4b57b67-2b57-4b96-be8b-807d25e2d8b4 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T03:12:39Z |
| publishDate | 2017 |
| publisher | Taylor and Francis |
| record_format | dspace |
| spelling | oxford-uuid:b4b57b67-2b57-4b96-be8b-807d25e2d8b42022-03-27T04:28:14ZMonoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasmaJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:b4b57b67-2b57-4b96-be8b-807d25e2d8b4EnglishSymplectic Elements at OxfordTaylor and Francis2017Lund, JElimar Bitsch, AGrønbech Rasch, MEnoksson, MTroeberg, LNagase, HLoftager, MOvergaard, MPetersen, HDecysin-1 (ADAMDEC1) is an orphan ADAM-like metalloprotease with unknown biological function and a short domain structure. ADAMDEC1 mRNA has previously been demonstrated primarily in macrophages and mature dendritic cells. Here, we generated monoclonal antibodies (mAbs) against the mature ADAMDEC1 protein, as well as mAbs specific for the ADAMDEC1 pro-form, enabling further investigations of the metalloprotease. The generated mAbs bind ADAMDEC1 with varying affinity and represent at least six different epitope bins. Binding of mAbs to one epitope bin in the C-terminal disintegrin-like domain efficiently reduces the proteolytic activity of ADAMDEC1. A unique mAb, also recognizing the disintegrin-like domain, stimulates the caseinolytic activity of ADAMDEC1 while having no significant effect on the proteolysis of carboxymethylated transferrin. Using two different mAbs binding the disintegrin-like domain, we developed a robust, quantitative sandwich ELISA and demonstrate secretion of mature ADAMDEC1 protein by primary human macrophages. Surprisingly, we also found ADAMDEC1 present in human plasma with an approximate concentration of 0.5 nM. The presence of ADAMDEC1 both in human plasma and in macrophage cell culture supernatant were biochemically validated using immunoprecipitation and Western blot analysis demonstrating that ADAMDEC1 is secreted in a mature form. |
| spellingShingle | Lund, J Elimar Bitsch, A Grønbech Rasch, M Enoksson, M Troeberg, L Nagase, H Loftager, M Overgaard, M Petersen, H Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title | Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title_full | Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title_fullStr | Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title_full_unstemmed | Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title_short | Monoclonal antibodies targeting the disintegrin-like domain of ADAMDEC1 modulates the proteolytic activity and enables quantification of ADAMDEC1 protein in human plasma |
| title_sort | monoclonal antibodies targeting the disintegrin like domain of adamdec1 modulates the proteolytic activity and enables quantification of adamdec1 protein in human plasma |
| work_keys_str_mv | AT lundj monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT elimarbitscha monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT grønbechraschm monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT enokssonm monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT troebergl monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT nagaseh monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT loftagerm monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT overgaardm monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma AT petersenh monoclonalantibodiestargetingthedisintegrinlikedomainofadamdec1modulatestheproteolyticactivityandenablesquantificationofadamdec1proteininhumanplasma |