KMID : 0545120130230040527
|
|
Journal of Microbiology and Biotechnology 2013 Volume.23 No. 4 p.527 ~ p.533
|
|
Production of Mouse Anti-Quail IgY and Subsequent Labeling with Horseradish Peroxidase Using Cyanuric Chloride
|
|
Neema Kassim
Adelard B. Mtenga Shim Won-Bo Chung Duck-Hwa
|
|
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 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.
|
|
KEYWORD
|
|
mouse anti-quail IgY, HRP labeling, cyanuric chloride, ELISA, SDS-PAGE
|
|
FullTexts / Linksout information
|
|
|
|
Listed journal information
|
|
|
|