Production of Mouse Anti-Quail IgY and Subsequent Labeling with Horseradish Peroxidase Using Cyanuric Chloride

• Published in: Journal of microbiology and biotechnology Vol. 23; no. 4; pp. 527 - 533
• Main Authors: Kassim, Neema, Mtenga, Adelard B, Shim, Won-Bo, Chung, Duck-Hwa
• Format: Journal Article
• Language: Korean
• Published: 한국미생물생명공학회 30.04.2013
• Subjects: cyanuric chloride; ELISA; HRP labeling; Mouse anti-quail IgY; SDS-PAGE; SDS-PAGE; HRP labeling; Mouse anti-quail IgY; cyanuric chloride; ELISA
• ISSN: 1017-7825; 1738-8872

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 105 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.