Single chain antibodies specific for fatty acids derived from a semi-synthetic phage display library

• Published in: Biochimica et biophysica acta Vol. 1569; no. 1; pp. 167 - 173
• Main Authors: Gargir, Ari, Ofek, Itzhak, Meron-Sudai, Shiri, Tanamy, Meital Gal, Kabouridis, Panagiotis S, Nissim, Ahuva
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.01.2002
• Subjects: Antibodies - immunology; Bacteriophage M13 - genetics; Bacteriophage M13 - immunology; Fatty Acids - immunology; Glycolipids - immunology; Gram-Positive Bacteria - immunology; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Lipoteichoic acid; Methyl palmitate; Palmitates - immunology; Peptide Library; Phage display; Single chain variable fragment; Phage display; LTA, lipoteichoic acid; Lipoteichoic acid; scFv, single chain variable fragment; TPBS, phosphate buffered saline containing 0.05% Tween 20; MP, methyl palmitate; PBS, phosphate buffered saline; Methyl palmitate; LPS, lipopolysaccharide; MPBS, skimmed milk in phosphate buffered saline; Single chain variable fragment
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/S0304-4165(01)00245-8

The biological activities of many acylated molecules are lipid dependent. Lipids, however, are poorly immunogenic or non-immunogenic. We employed a phage display semi-synthetic human antibody library to isolate anti-lipid antibodies. Selection was done against methyl palmitate, a 16 carbon aliphatic chain, and a major component of bacterial glycolipids and lipoproteins in animal cells. The selected single chain variable fragment (scFv) bound specifically to a 16 carbon aliphatic chain and to a lesser extent to a 14 or 18 carbon aliphatic chain and poorly to either 12, 22 or 8 carbon aliphatic chains. Furthermore, the scFv prevented micelle formation of lipoteichoic acid from Gram-positive bacteria; inhibited lipopolysaccharide-induced tumor necrosis factor α release in mononuclear cells; bound to hydrophobic bacterial surfaces, especially those of Gram-positive bacteria, and bound to Lck, a mammalian palmitated lipoprotein. Our data suggest that the phage antibody library can be successfully employed to obtain human anti-aliphatic scFv human antibody fragment with potential therapeutic applications in neutralizing the deleterious effects of bacterial toxins as well as in structure–function analysis of lipoproteins in animal cells.