4.4.2. Serum Preparation

Two 3-month old female New Zealand white rabbits were immunized intradermally with the CONA-hapten conjugate (immunogen). Rabbit immunisation was performed under the supervision of the Ethics Committee of Research on Animals (Food Science Research Institute, Institute of Food Sciences, Hungarian University of Agriculture and Life Sciences, Budapest, Hungary) and under the authorisation and inspection by the Government Office for Pest County in Hungary (Official permit for animal testing # PE/EA/45-6/2020, last date of approval: 21 February 2020). Initial immunization was carried out with 0.1 mg immunogen dissolved in PBS and emulsified in Freund's complete adjuvant (1:1 volume fraction). Then, injection of 0.15 mg of the immunogen in PBS and Freund's incomplete adjuvant (1:1 volume fraction) was given (the first three booster injections at 3-week intervals, with the subsequent one at a 1-month interval). One week after each immunization, rabbits were bled and after coagulation of the blood (4 ◦C overnight) the sera were centrifuged (2400 g, 15 min) and purified by gel chromatography on PD-10 desalting columns.
