Development Of An Enzyme-Linked Immunosorbent Assay For The Detection Of Salbutamol

The hapten salbutamol succinate derivative was prepared from salbutamol sulphate.The hapten was coupled to KLH for use as the immunogen SAL-KLH, and coupled to HRP for use as the enzyme tracer with method of mixed acid anhydride. The hapten SAL was coupled to the protein OVA with method of carbodiimide to be used as coating conjugate for indirect competitive ELISA.The antiserum obtained from Japan big rabbit immunised by SAL-KLH showed high affinity to SAL. A rapid indirect competitive enzyme-linked immunosorbent assay (ELISA) procedure employing a polyclonal antibody purified by immunity affinity chromatography from the antiserum was established. The antibody showed high sensitivity and specificity in phosphate buffer, with an IC50of1.7ng/mL, and the limit of detection was0.09ng/mL. Besides, the antibody showed no cross-reactivity with ractopamine, dobutamine and isoxsuprine. Choose pork to the elimination of matrix effects study. When the concentration of salbutamol at a higher level, the substrate curves coincide well with the standard curve, when the concentration of salbutamol at a low level, the matrix curve does not coincide with the standard curve. The recoveries of SAL was more than70%at three different levels. It is a basic of SAL-ELISA kit development.In addition, the stored condition of SAL-ELISA kit was studied to validate the stabiblity of antibody and enzyme tracer. The results showed that it could be stored at4℃over half a year without any decline in detect capability. It benefits for the SAL-ELISA kit’s application and important to control the use of salbutamol.