Preparation and qualification of estradiol antigen and antiserum

In order to develop enzyme immunoassay for estradiol,the antigen and antiserum of estradiol were prepared.Estradiol was coupled to bovine serum albumin(BSA) and ovalbumin(OVA) by using the ester activation method.The conjugates were identified by UV absorption method to calculate the coupling ratio and the protein concentration.E2-BSA was used as immunogen for injection of rabbits to prepare antiserum.The affinity and titer of antiserum were detected by using the E2-OVA as coating antigen.The results indicated that estradiol antigen was synthesized successfully.The protein concentration of E2 to BSA was 5.455 mg/mL and to OVA was 7.533 mg/mL.The coupling ratio of E2 to BSA was 7:1 and to OVA was 8:1.The titer of antiserum was 1:106.