Asian Journal of Nuclear Medicine
亞洲核醫學雜志

Volume 1 Number 1, April 2001
Abstract

AFP時間分辨熒光免疫分析方法學研究

黃C、蔡剛明、肖華龍、朱利國、 譚成、金堅
江蘇省原子醫學研究所,核醫學國家重點實驗室,無錫 214063,中國

目的 建立AFP時間分辨熒光免疫分析 方法 以AFP單克隆抗體A47包被板,雙功能螯合劑異氰酸R基二乙烯三胺四乙酸絡合Eu3+及標記A137單抗,發光增強系統為以β-二酮體為主的增強液。採用平衡飽和法建立AFP時間分辨熒光免疫分析,資料採用Log-Logit法函數和四參數Logitc函數資料處理程式處理。結果 方法的批內和批間CV分別為 1.59%和7.14%,平均回收率 為103.98%,靈敏度為 0.43μg/L,可測範圍為 4.22-1210μg/L,ED20、ED50和ED80分別為 4.248μg/L、35.59μg/L和246.9μg/L。本方法與CEA、CA125、CA153和CA199均無交叉反應。Eu3+標記抗體-30℃保存一年多免疫反應性基本無損失,同批試劑連續5個月應用分析結果穩定。與Wallac同類產品比較,結果吻合,表明國產AFP單抗可取代進口,AFP時間分辨熒光免疫分析的質量參數優於EIA,RIA和IRMA。

【關鍵字】 時間分辨熒光免疫分析;甲胎蛋白

Study of time-resolved fluoroimmunoassay of α-fetoprotein

HUANG Biao, CAI Gangmin, XIAO Hualong, ZHU Liguo, TAN Cheng, JIN Jian. Jiangsu Institute of nuclear medicine, State Key Laboratory of Nuclear Medicine, Wuxi 214063,China.

Objective To set a two-site time-resolved fluoroimmunoassay (TRFIA) of α-fetoprotein (AFP) .Methods A two-siteTRFIA of AFP based on the direct sandwich technique has been developed, on the equilibrium method. The monoclonal antibody (MoAb) directed against a specific antigenic site on the AFP was A47. The europium-labelled MoAb of A137, which was prepared using N- (p-isothiocyanatobenzyl)-diethylenetriamine-N, N, N, N-tetraacetic acid, was directed against a different antigenic site on AFP. The luminescent enhancement system was a solution mainly containing 2-naphthoyltrifluoroacetone. Results The intra- and inter- assay CV of the TRFIA were 1.59% and 7.14%, respectively, and the recovery rate was 103.98%, the sensitivity was 0.43μg/L. The MoAb provided a linear response from 4.22 to 1210μg/L with ED50 of 35.59μg/L. The cross- reacting rate of this study with CEA, CA125, CA199 and CA153 measurements was negligible. The europium-labelled A137 was stable for a year at -30℃ and the results of the TRFIA with same reagents were stable in five months. Same results were observed when compared to product of Wallac Co. Conclusion : The A137 and A47 were satisfactory for TRFIA, and the parameters of this assay were better than of the EIA's, RIA's and IRMA's. (Asian J Nucl Med 2001;1:40-42)

Key Words : time-resolved fluoroimmunoassay, α-fetoprotein.



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