Development of a monoclonal antibody that specifically detects tissue inhibitor of metalloproteinase-4 (TIMP-4) in formalin-fixed, paraffin-embedded human tissues.
Overexpression of the extracellular metalloproteinase inhibitor TIMP-4 in estrogen receptor-negative breast cancers was found recently to be associated with a poor prognosis for survival. To pursue exploration of the theranostic applications of TIMP-4, specific antibodies with favorable properties for immunohistochemical use and other clinical assays are needed. Here we report the characterization of a monoclonal antibody (clone 9:4-7) specific for full-length human TIMP-4 with suitable qualities. The antibody was determined to be an IgG(2b) immunoglobulin. In enzyme-linked immunosorbent assay (ELISA) and immunoblotting assays, it did not exhibit any detectable crossreactivity with recombinant forms of the other human TIMPs 1, 2, and 3. In contrast, the antibody displayed high specificity and sensitivity for TIMP-4 including in formalin-fixed and paraffin-embedded specimens of human breast specimens. An analysis of tissue microarrays of human cancer and corresponding normal tissues revealed specific staining patterns with excellent signal-to-noise ratios. This study documents TIMP-4 monoclonal antibody clone 9:4-7 as an effective tool for preclinical and clinical investigations.
Donover
P Scott
PS
Lankenau Institute for Medical Research, Wynnewood, Pennsylvania 19096, USA.
Wojciechowski
Brian S
BS
Thirumaran
Rajesh
R
Zemba-Palko
Vlasta
V
Prendergast
George C
GC
Wallon
U Margaretha
UM
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007150:Immunohistochemistry / Q000379:methods; D016612:Paraffin Embedding; D046888:Tissue Array Analysis / Q000379:methods*; D019714:Tissue Inhibitor of Metalloproteinases / Q000276:immunology*; D000097792:Tissue Inhibitor of Metalloproteinase-4
D000911:Antibodies, Monoclonal; D005404:Fixatives; D005557:Formaldehyde; D007074:Immunoglobulin G; D019714:Tissue Inhibitor of Metalloproteinases; D000097792:Tissue Inhibitor of Metalloproteinase-4