發(fā)布時間：2018-07-03 07:38

  本文選題：BHHCT + BBCAP��； 參考：《華中科技大學(xué)》2012年碩士論文

【摘要】：作為一種新型免疫分析方法，時間分辨熒光免疫分析法(TRFIA)選用稀土絡(luò)合物作為標(biāo)記物，這類標(biāo)記物熒光壽命長、Stokes位移大、特征峰尖銳，因此可以通過延遲測量時間的方法，有效地消除背景熒光的干擾，從而具備了相比一般免疫分析方法更高的靈敏度。 本工作的主要目的是建立基于AP酶的ELISA和熒光酶免疫分析(FEIA)，以及時間分辨熒光免疫分析方法，其中涉及到稀土螯合物BBCAP、BHHCT和抗體的偶聯(lián)，以及使用時間分辨熒光免疫分析方法進(jìn)行免疫球蛋白抗體的測定。 本論文研究的主要內(nèi)容有： （1）以BBCAP作為配體，與羊抗小鼠IgG偶聯(lián)，測定其免疫特性。 （2）合成BHHCT-羊抗小鼠IgG偶聯(lián)物，并對Eu~(3+)標(biāo)記抗體技術(shù)進(jìn)行了實驗研究，討論了Eu~(3+)濃度、稀釋后的偶聯(lián)物與Eu~(3+)使用液混合后放置時間、Eu~(3+)放置時間、偶聯(lián)物稀釋倍數(shù)、光致漂白效應(yīng)、溫度、Wallac洗液對免疫反應(yīng)的影響。優(yōu)化后采取的實驗條件為：Eu~(3+)使用濃度為1*10-5M，偶聯(lián)物稀釋倍數(shù)為20倍、偶聯(lián)物與Eu~(3+)使用液混合后放置時間為60min。 （3）以小鼠IgG為抗原，羊抗小鼠IgG為抗體，建立了競爭酶聯(lián)免疫吸附測定（ELISA）、熒光酶免疫分析（FEIA）和時間分辨熒光免疫分析（TRFIA）體系，ELISA、FEIA、TRFIA達(dá)到的靈敏度分別為IC(20)=2.477μg/mL，IC(50)=21.115μg/mL；IC(20)=2.522μg/mL，， IC(50)=30.723μg/mL； IC(20)=0.113μ g/mL， IC(50)=1.131μg/mL。
[Abstract]:As a new immunoassay method, time resolved fluorescence immunoassay (TRFIA) uses rare earth complexes as markers. The fluorescence lifetime of these markers has long Stokes shift and sharp characteristic peaks, so they can be measured by the method of delayed measurement of time. The interference of background fluorescence is eliminated effectively and the sensitivity is higher than that of general immunoassay. The main purpose of this work is to establish an AP enzyme based Elisa and fluorescence enzyme immunoassay (FEIA), and a time resolved fluorescence immunoassay method involving the coupling of rare earth chelates BBCAPHHCT and antibodies. Time-resolved fluorescence immunoassay was used to detect the immunoglobulin antibody. The main contents of this thesis are as follows: (1) using BBCAP as ligand, coupling with goat anti-mouse IgG to determine its immunological characteristics. (2) synthesizing BHHCT- goat anti-mouse IgG conjugate, and studying the technique of Eu3 labeled antibody. The effects of EU ~ (3) concentration, the time of depositing EU ~ (3) after mixed with EU ~ (3), dilution multiple, photobleaching effect and temperature Wallac lotion on the immune reaction were discussed. The optimized experimental conditions were as follows: the concentration of 10 ~ (-3) was 1 ~ 10 ~ (-5) M, the dilution multiple of the conjugate was 20 times, and the time of storage was 60 min after mixing the conjugate with EU ~ (3). (3) the mouse IgG was used as antigen and sheep anti-mouse IgG as antibody. Competitive enzyme linked immunosorbent assay (Elisa), fluorescence enzyme immunoassay (FEIA) and time resolved fluorescence immunoassay (TRFIA) were established. The sensitivity of ELISAA FEIAA TRFIA was 2.477 渭 g / mL IC (50) 21.115 渭 g / mL IC (20) 2.522 渭 g / mL, IC (50) 30.723 渭 g / mL; IC (20) 0.113 渭 g / mL, IC (50) 1.131 渭 g / mL.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R392.1

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前2條

1 海曉丹;時間分辨熒光免疫分析新方法研究[D];中國科學(xué)院研究生院（大連化學(xué)物理研究所）;2004年

2 吳麗;免疫球蛋白的時間分辨熒光免疫分析方法研究[D];華中科技大學(xué);2008年

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