Cat. No.
MABL-2381
Application
functional assays, in vivo, SPR, ELISA
Isotype
Engineer antibody
Species Reactivity
Human, Mouse
Clone No.
Nb14
From
Recombinant Antibody
Specificity
This antibody is specific for mouse MMP8. It was found to be cross-reactive with the catalytic domain of human MMP8 (hMMP8_CD). This antibody has inhibitory capacity, achieved by steric hindrance of MMP8 or induction of conformational changes in MMP8 rather than direct binding to the catalytic pocket. This antibody does not exhibit cross-reactivity with other MMPs (human) and ADAM10/ADAM17.
Alternative Names
MMP-8; Neutrophil collagenase; Matrix metalloproteinase-8; Matrix Metallo-Proteinase 8; Collagenase 2; PMNL-CL; PMNL collagenase; MNL-CL; CLG1; HNC; Nb 70; Nanobody 70
UniProt
O70138
Immunogen
The original antibody was generated by immunizing an alpaca (Vicugna pacos) with the recombinant mouse catalytic domain of matrix metalloproteinase 8 (mMMP8_CD). A VHH library was constructed using total RNA from peripheral blood lymphocytes, and phages displaying the VHH repertoire were screened for mMMP8_CD-specific nanobodies.
Application Notes
The binding affinity of this antibody's original format (VHH) to different substrates, including the catalytic domain of mouse MMP8 (mMMP8_CD), full-length mouse MMP8 (mMMP8_FL), and the catalytic domain human MMP8 (hMMP8_FL), was measured by ELISA. It had a KD of 0.24 nM with mMMP8_CD, a KD of 1.69 nM with mMMP8_FL, and a KD of 158.4 nM with hMMP8_CD. SPR analysis was conducted to measure its binding kinetics to mMMP8_CD; a KD of 1.334 ± 0.137 nM was measured. Its in vivo pharmacokinetic properties were studied by injecting a single dose intraperitoneally into mice; blood samples were collected at different time points, and the nanobody's level in the mouse serum was detected using ELISA to evaluate its persistence and clearance in the bloodstream over time.
Antibody First Published
Demeestere et al. Development and Validation of a Small Single-domain Antibody That Effectively Inhibits Matrix Metalloproteinase 8 Mol Ther. 2016 May;24(5):890-902. doi: 10.1038/mt.2016.2 PMID:26775809
Note on publication
The original publication focuses on the development and validation of a small single- domain antibody (nanobody) that effectively inhibits Matrix Metalloproteinase 8 (MMP8).
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
