[Preparation of monoclonal antibody against sulfadiazine and development of an ELISA kit].
OBJECTIVE
To prepare monoclonal antibody (mAb) against sulfadiazine (SD) and develop an ELISA kit for rapidly detecting residual SD in different kinds of samples.


METHODS
BALB/c mice were immunized with the conjugate of SD and BSA, and then anti-SD mAb was prepared by hybridoma technique. The purified ascitic mAb and HRP-labeled SD were used to establish a competitive ELISA for detection of SD in samples.


RESULTS
5 hybridoma cell lines secreting anti-SD mAbs 1A1, 1B8, 1E4, 2C1 and 3D9 were obtained. 1A1, 1B8 and 1E4 belonged to IgG1, while 2C1 and 3D9 belonged to IgG2. The I50 and theoretical minimum detectable amount of the kit was 9.3 microg/L and 0.6 microg/L, respectively. The recovery rates of the kit for SD in different kinds of samples were higher than 60%. The cross-reaction rate of the kit for other sulfanilamide drugs was lower than 3%.


CONCLUSIONS
5 mAbs against SD have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for rapidly detecting SD can meet the needs of detection of SD in different samples.
Zhao
Pi-cheng
PC
Laboratory of Cytology and Immunology, Shanghai Cygen (Shanghai) Biotech Co., Ltd., Shanghai 200233, China. Cygen@sh163.com
Yuan
Da-wei
DW
Zhu
Run
R
Ji
Jing-hui
JH
Zhu
Hong-wei
HW
Zhang
Xiao-feng
XF
Qiu
Lie-qun
LQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011933:Reagent Kits, Diagnostic; D013411:Sulfadiazine / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D011933:Reagent Kits, Diagnostic; D013411:Sulfadiazine