Process for the production of a hybridoma and antibody obtained therefrom, able to recognize more than one vitamin D metabolite
Inventors
Anciaux, Michel • Mathieu, Fabienne • Lin, Frederic • Poncelet, Martin
Assignees
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Publication Number
US-9746483-B2
Publication Date
2017-08-29
Expiration Date
Abstract
The invention concerns a process for the production of a hybridoma, and of a monoclonal antibody or fragments thereof able to recognize 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3.
Core Innovation
The invention relates to monoclonal antibodies and hybridomas for recognizing 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3. Monoclonal antibodies derived from a single hybridoma raised using one hapten of general formula (I) bind both 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3.
The invention addresses the need to recognize more than one vitamin D metabolite using monoclonal antibodies derived from the hybridomas. It links hapten/conjugate design to immunogen preparation, including activation/coupling via the carboxyl function and carrier proteins, and describes producing antibodies using a hybridoma workflow involving immunization and B-cell fusion with myeloma.
The document reports binding/inhibition and simultaneous-recognition performance, including in vitro tests, coated-tube immunoassays, and validation using LC-MS in human serum. It also extends to diagnostic devices/kits and signal tracers/biosensor modalities using vitamin D-related tracer formulae and labeling groups such as 125I, enzymes such as HRP/alkaline phosphatase, and related tracer constructs.
Claims Coverage
The provided independent claims cover two inventive features: specific hybridomas suitable for producing a monoclonal antibody recognizing both 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3, and monoclonal antibodies produced from those specific hybridomas, as defined by BCCM/LMBP deposit numbers.
Hybridoma deposit selection for dual metabolite recognition
A hybridoma suitable for the production of a monoclonal antibody able to recognize 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3, selected from the hybridomas deposited in the BCCM/LMBP under deposit numbers LMBP 7011CB, LMBP 7012CB, LMBP 7013CB, LMBP 7204CB and LMBP 7205CB.
Monoclonal antibody produced from specified hybridoma deposits
A monoclonal antibody produced from a hybridoma selected from the hybridomas deposited in the BCCM/LMBP under deposit numbers LMBP 7011CB, LMBP 7012CB, LMBP 7013CB, LMBP 7204CB and LMBP 7205CB.
Overall, the claim coverage is centered on monoclonal antibody recognition of both 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3, with the defining inventive limitation being that the antibodies are produced from hybridomas selected by specific BCCM/LMBP deposit numbers.
Stated Advantages
Monoclonal antibodies can recognize both 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3.
Simultaneous-recognition performance is reported, including binding/inhibition results and validation using LC-MS in human serum.
Documented Applications
Diagnostic devices/kits are described for vitamin D-related detection using monoclonal antibodies, including tracer/signal modalities and biosensor formats.
In vitro tests and coated-tube immunoassays are described, with validation using LC-MS in human serum.
Signal tracers/biosensor modalities are described using vitamin D-related tracer formulae and labeling groups such as 125I and enzymes such as HRP/alkaline phosphatase.
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