title: Production of mouse anti-quail IgY and subsequent labeling with horseradish peroxidase using cyanuric chloride.
abstract: Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was 10(5) CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.
authors.affiliation: Division of Applied Life Science (BK 21 program), Gyeongsang National University, Jinju Gyeongnam 660 - 701, Korea.
authors.forename: Neema
authors.initials: N
authors.lastname: Kassim
authors.forename: Adelard B
authors.initials: AB
authors.lastname: Mtenga
authors.forename: Won-Bo
authors.initials: WB
authors.lastname: Shim
authors.forename: Duck-Hwa
authors.initials: DH
authors.lastname: Chung
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001431:Bacteriological Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006041:Goats; D006735:Horseradish Peroxidase / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D007136:Immunoglobulins / Q000276:immunology; D051379:Mice; D011784:Quail / Q000276:immunology; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D014227:Triazines / Q000378:metabolism; D014736:Vibrio parahaemolyticus / Q000302:isolation & purification*; D041261:Vibrio vulnificus / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; C016512:IgY; D007074:Immunoglobulin G; D007136:Immunoglobulins; D014227:Triazines; C019309:cyanuric chloride; D006735:Horseradish Peroxidase