Research on Preparation of Artificial Antigen of Procymidone

doi:10.11924/j.issn.1000-6850.casb2021-0017

Abstract

Abstract:

To prepare the artificial antigen of procymidone, the succinic anhydride method was applied to modify the structure of procymidone, and the modified procymidone was coupled with bovine serum albumin (BSA) and ovalbumin (OVA) by 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC) method to synthetize PCM-BSA and PCM-OVA. SDS-PAGE, UV scanning and animal immunization were used to identify the quality of artificial antigen of PCM. The results of SDS-PAGE and UV scanning showed that PCM was coupled with the carrier proteins successfully. The serum of mice immunized by PCM-BSA could react with PCM-OVA; the serum titer could be 1:6400. The PCM was applied as an inhibitor to prevent the reaction between serum and PCM-OVA. However, even the concentration of PCM reached to 256 µg/mL, there was no significant inhibition. When PCM was replaced by the prepared PCM-OVA solution, which could inhibit the reaction between serum and PCM-OVA coated on the microtiter plates, the 50% inhibition concentration (IC₅₀) was 320 times diluted of the prepared PCM-OVA solution. In this study, PCM was coupled with carrier proteins successfully, but the serum sensitivity of mouse immunized with PCM artificial antigen was not high, which indicated that the benzene ring might be part of antigenic determinant of PCM. The epitope of PCM was destroyed during molecular modification by succinic anhydride, so this method was not suitable for the preparation of PCM artificial antigen.

Key words: procymidone, structural modification, artificial antigen, animal immunization, serum

CLC Number:

R155.5

Hu Xiaofei, Xing Yunrui, Sun Yaning, Pang Xinghao, Wang Yao. Research on Preparation of Artificial Antigen of Procymidone[J]. Chinese Agricultural Science Bulletin, 2021, 37(33): 142-147.

Figures/Tables 6

References 27

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项目		血清稀释倍数							空白
-	小鼠序号	1:200	1:400	1:800	1:1600	1:3200	1:6400	1:12800	空白
OD₄₅₀值	1	2.005	1.336	0.821	0.599	0.375	0.209	0.143	0.079
	2	1.511	0.723	0.430	0.311	0.358	0.097	0.060	0.065
	3	0.808	0.361	0.207	0.140	0.103	0.073	0.076	0.068

项目		血清稀释倍数							空白
-	小鼠序号	1:200	1:400	1:800	1:1600	1:3200	1:6400	1:12800	空白
OD₄₅₀值	1	2.005	1.336	0.821	0.599	0.375	0.209	0.143	0.079
	2	1.511	0.723	0.430	0.311	0.358	0.097	0.060	0.065
	3	0.808	0.361	0.207	0.140	0.103	0.073	0.076	0.068

项目		PCM浓度/(µg/mL)								阳性	空白
-	序号	256	128	64	32	16	8	4	2	阳性	空白
OD₄₅₀值	1	0.714	0.759	1.003	0.911	0.850	0.926	0.755	0.781	0.792	0.087
	2	0.838	0.873	0.932	0.911	0.912	0.941	0.840	0.845	0.864	0.053
	3	0.546	0.693	0.799	0.827	0.601	0.666	0.656	0.606	0.544	0.064

项目		PCM浓度/(µg/mL)								阳性	空白
-	序号	256	128	64	32	16	8	4	2	阳性	空白
OD₄₅₀值	1	0.714	0.759	1.003	0.911	0.850	0.926	0.755	0.781	0.792	0.087
	2	0.838	0.873	0.932	0.911	0.912	0.941	0.840	0.845	0.864	0.053
	3	0.546	0.693	0.799	0.827	0.601	0.666	0.656	0.606	0.544	0.064

项目		PCM-OVA稀释倍数						阳性	空白
-	序号	10	20	40	80	160	320	阳性	空白
OD₄₅₀值	1	0.289	0.390	0.477	0.601	0.700	0.813	1.514	0.051
	2	0.163	0.151	0.235	0.245	0.282	0.341	0.533	0.058
	3	0.209	0.308	0.454	0.543	0.677	0.686	1.352	0.052