Generation and characterization of chicken monoclonal antibodies  against human LOX-1.
Lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1) is the major receptor for oxidized LDL (oxLDL), and plays a key role in the pathogenesis of atherosclerosis and cardiovascular diseases. Monoclonal antibodies (mAbs) specific for human LOX-1 (hLOX-1) were generated by a phage display technique using chickens immunized with recombinant hLOX-1 (rhLOX-1). A total of 53 independent scFv clones reactive for rhLOX-1 were obtained. Of the 53 clones, 49 recognized the C-type lectin-like domain (CTL domain), which contributes to the binding of oxLDL. Of these, 45 clones inhibited oxLDL-binding with LOX-1. Furthermore, some of these clones cross-reacted with rabbit, pig and/or mouse LOX-1. For possible application as therapeutic agents in the future, two cross-reactive mAbs were re-constructed as chicken-human chimeric antibodies. The chimeric antibodies showed similar characteristics compared to the original antibodies, and inhibited oxLDL binding to LOX-1 expressed on CHO cells. The results obtained in this study indicate that anti-LOX-1 mAbs might be useful tools for functional analyses and development of therapeutic agents for cardiovascular indications such as atherosclerosis.
Iwamoto
Shin
S
Laboratory of Immunobiology, Department of Molecular and Applied Bioscience, Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan.
Nishimichi
Norihisa
N
Tateishi
Yoshiko
Y
Sato
Yuko
Y
Horiuchi
Hiroyuki
H
Furusawa
Shuichi
S
Sawamura
Tatsuya
T
Matsuda
Haruo
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D051379:Mice; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics; D051127:Scavenger Receptors, Class E / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology*; D013552:Swine
D000911:Antibodies, Monoclonal; C495137:OLR1 protein, human; D011994:Recombinant Proteins; D051127:Scavenger Receptors, Class E