title: Development and characterization of a mouse monoclonal antibody against antimicrobial peptide tachyplesin I.
abstract: Monoclonal antibodies against tachyplesin I (TP I) were developed to study its mechanisms of activity, a kind of cationic antimicrobial peptides (AMPs), in vivo or in vitro, and to purify TP I from expression products. The synthesized TP I was chemically conjugated with the carrier protein BSA and then injected into BALB/c mice. Positive hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using TP I and subcloned three times with limiting dilution. Five MAbs effective in detecting the native TP I (named 2D8, 3B8, 5H2, 6B12, and 8F5) were obtained. Isotyping of all obtained MAbs indicated that MAbs 2D8, 3B8, 5H2, and 8F5 belong to IgG1, and MAb 6B12 belongs to IgG2a. Specificity assay showed that MAb 8F5 had almost the same level of specificity to natural TP I, recombinant TP I, and synthesized TP I and TP II, but did not cross-react with control peptides. These results suggest that the synthetic AMP conjugates can elicit antibodies against native AMPs and can be used to detect antimicrobial peptides.
authors.affiliation: Animal Science and Veterinary College, Jilin University, 5333 Xi'an Road, Changchun, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Feng
authors.forename: Wenyu
authors.initials: W
authors.lastname: Han
authors.forename: Zhanyun
authors.initials: Z
authors.lastname: Song
authors.forename: Honglei
authors.initials: H
authors.lastname: Zhao
authors.forename: Yu
authors.initials: Y
authors.lastname: Gao
authors.forename: Yuwen
authors.initials: Y
authors.lastname: Diao
authors.forename: Shanshan
authors.initials: S
authors.lastname: Liu
authors.forename: Liancheng
authors.initials: L
authors.lastname: Lei
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry / Q000378:metabolism*; D000918:Antibody Specificity; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D010456:Peptides, Cyclic / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D017186:Titrimetry
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D023181:Antimicrobial Cationic Peptides; D004268:DNA-Binding Proteins; D007074:Immunoglobulin G; D010446:Peptide Fragments; D010456:Peptides, Cyclic; C057636:tachyplesin peptide, Tachypleus tridentatus; D012710:Serum Albumin, Bovine