Mfg Part Number
Ab04110-10.159

Vector Labs Ab04110-10.159 Anti-MMP8 [Nb14], Human IgG1-Fc Fusion, (100 μg)

Quick Overview
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.
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Product Class:Purified
Clone ID:Nb14
Synonyms: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
Amount:100 μg
Application Codes Clone:ELISA; SPR; functional assays; in vivo
Shipping Temperature:Wet Ice
Origin Pub PMID:26775809
Buffer Composition:PBS with 0.02% Proclin 300.
Original Format:VHH
Storage Temperature:Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at -20⁰C.
Chimeric Use Statement:This chimeric human antibody was made using the variable domain sequences of the original Alpaca VHH format, for improved compatibility with existing reagents, assays and techniques.
Specificity Statement: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.
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. This antibody's inhibitory capacity against MMP8 was investigated; it demonstrated the strongest inhibition of mMMP8_CD with an IC50 of 4.359 µM using DQ gelatin as a substrate and an IC50 value of 19.5 µM using DQ collagen type I. This antibody was used to create a bivalent nanobody by linking it to itself and to clone Nb Albumin using flexible [G4-S]3 linkers. The resulting nanobodies (Nb 14-Alb and Nb 14-Alb-14) were used in in vivo experiments for various applications (Demeestere et al., 2016; PMID: 26775809). The original format of this antibody was used to create another trivalent nanobody by linking it to clones Nb 70 and Nb Albumin using the same linkers. The resulting nanobody (Nb 70-Alb-14) was used in vivo to study the effects of inhibiting matrix MMP8 and TNFR1 on sepsis, using endotoxemia and CLP (cecal ligation and puncture; a surgical procedure commonly used in animal models to induce polymicrobial sepsis) experimental models (Steeland et al., 2018; PMID: 29095202).
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Brand Vector Labs
UNSPSC 12161500