Preparation of artificial antigen and egg yolk-derived immunoglobulin (IgY) of citrinin for enzyme-linked immunosorbent assay.
OBJECTIVE
To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin (IgY) to build an enzyme-linked immunosorbent assay (ELISA) for citrinin (CTN).


METHODS
CTN was conjugated with bovine serum albumin (BSA), ovalbumin (OVA) with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet (UV) spectrometry and Infrared (IR) spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied.


RESULTS
UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng x mL(-1), with a good linearity ranging 20-640 ng x mL(-1).


CONCLUSIONS
Artificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.
Duan
Zhao-Hui
ZH
Laboratory of Clinical Immunology, Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510120, Guangdong, China. China-1@Tom.com
Lin
Zhuang-Sen
ZS
Yao
He-Rui
HR
Gao
Yan-Hong
YH
Zhang
Kun
K
Zhao
Su-Qing
SQ
Zhu
Zhen-Yu
ZY
D000818:Animals; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry*; D002645:Chickens; D002953:Citrinin / Q000737:chemistry*; D004530:Egg Yolk / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D007136:Immunoglobulins / Q000276:immunology*
D000941:Antigens; C016512:IgY; D007136:Immunoglobulins; D002953:Citrinin