Properties of blocking and non-blocking monoclonal antibodies specific for human macrophage galactose-type C-type lectin (MGL/ClecSF10A/CD301).
Monoclonal antibodies (mAbs) specific for the human macrophage galactose-type calcium-type lectin (MGL) were established. The recombinant extracellular domain of MGL was used to immunize a mouse, and 10 hybridoma clones were obtained. Binding of recombinant MGL to asialo-bovine submaxillary mucin was shown to be blocked by mAbs MLD-1, 4 and 6. Immunoprecipitation of MGL from lysates of COS-1 cells transfected with MGL cDNA (form 6A) was achieved with mAbs MLD-1, 4, 7, 8 and 16. Chimeric recombinant proteins between human MGL and mouse MGL1 were used to determine the location of the epitopes for these mAbs. mAbs MLD-8, 13, 15 and 16 interacted with the amino terminal side of the conserved WVDGTD sequence immediately upstream of QPD, whereas mAbs MLD-7, 12 and 17 interacted with the other side. mAbs MLD-1, 4, and 6 apparently required both sides of this boundary. mAbs MLD-15 and 16 were shown to recognize the protein products of alternatively spliced mRNA 6A/8A and 6C/8A, having deletions at the boundary of exons 7 and 8, in addition to full length and other spliced forms of MGL (6A, 6B and 6C), whereas the other mAbs bound only full length and forms 6A, 6B and 6C.
Sano
Yoshihiko
Y
Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0003, Japan.
Usami
Katsuaki
K
Izawa
Ryota
R
Denda-Nagai
Kaori
K
Higashi
Nobuaki
N
Kimura
Toshifumi
T
Suzuki
Noriko
N
Irimura
Tatsuro
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001212:Asialoglycoproteins / Q000276:immunology; D015153:Blotting, Western; D019556:COS Cells; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002522:Chlorocebus aethiops; D000939:Epitopes / Q000187:drug effects / Q000276:immunology; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D047468:Immunoprecipitation; D037181:Lectins, C-Type / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D009077:Mucins / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D020298:U937 Cells
D000911:Antibodies, Monoclonal; D001212:Asialoglycoproteins; D000939:Epitopes; D037181:Lectins, C-Type; C506225:MGL lectin, human; D009077:Mucins; D011993:Recombinant Fusion Proteins; D002118:Calcium