RESUMO
Under low temperature conditions, the hapten carboxyl-norketamine was synthesized by reacting norketamine and succinaldehyde acid.Identification result using electrospray ionization mass showed the hap ten was successfully synthesized.The artificial antigen confirmed by infrared spectroscopy was developed by conjugating hapten to carrier proteins with carbodiimide(EDC) method.Matrix-assisted laser desorption ioni zation time of flight mass spectrometry(MALDI-TOF-MS) showed that the ratio of hapten to BSA was 11:1.The antibody with high titer(5.12 × 10~4) was produced after immuning to rabbits.
RESUMO
Preparation of an antibody-colloidal gold probe (conjugate) specific to aflatoxin B1 (AFB1) and its use in developing a rapid AFB1 diagnostic method was presented in this paper. Monodispersional nanogold colloid was synthesized and preparation of nanogold-labeled polyclonal antibody probe to aflatoxin B1 under friendly and optimal condition. Combination of antibody with nanogold particles was also characterized by UV-visible (UV-vis) light absorption spectra, transmission electron microscopy (TEM), fluorescence spectroscopy, titers, cross reactivity and stability measurements. Furthermore, nanogold-labeled probe was used to develop an immunochromatographic (IC) method for aflatoxin B1 analysis. With this method, analysis could be completed in less than 10 min. Detection time was reduced 6-10 times comparative with ELISA. With visual observation, lower test limit was found to be around 2.5 ng/ml aflatoxin B1 standard solution, which was increased to two times of ELISA.
