發(fā)布時間：2018-06-13 19:20

  本文選題：SPR生物傳感器 + IAA　； 參考：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：3-吲哚乙酸(IAA)和脫落酸(ABA)是普遍存在的植物激素,在植物生長發(fā)育過程中起著重要的作用。IAA最基本的功能不但可以加速和調(diào)控植物的生長發(fā)育,而且還可以阻礙植物的生長和根、莖、組織建成的作用。ABA的基本作用是促進和抑制植物生長,促進葉子與果實的脫落,影響花期等作用。目前這兩種植物激素的檢測方法大都存在樣品前處理復(fù)雜,費時,成本高,重現(xiàn)性和選擇性差等缺點。因此,本研究采用了具有實時監(jiān)測、樣品無需預(yù)處理、快速靈敏檢測等優(yōu)點的表面等離子體共振(SPR)生物傳感器檢測IAA和ABA這兩種植物激素。為了提高SPR傳感器的靈敏度,本文采用兩種方法:一種是將金/銀合金納米粒子作為敏化材料引入SPR生物傳感器中,另一種是通過加長檢測鏈的方式來提高SPR生物傳感器檢測的靈敏度。本文包括以下兩部分實驗內(nèi)容:(1)兩種SPR生物傳感器的構(gòu)建以及對IAA的檢測:一種是傳統(tǒng)模式SPR生物傳感器(傳感器1),另一種是引入Au/Ag合金納米粒子作為增敏材料的SPR生物傳感器(傳感器2)。首先用不同的物質(zhì)修飾SPR的芯片。傳感器1:先在裸金片上修飾MPA,再修飾蛋白A,蛋白A能與MPA末端的被EDC/NHS活化后的羧基結(jié)合使蛋白A修飾到芯片上,用鹽酸乙醇胺對芯片進行滅活,IAA抗體的非抗原結(jié)合位點能與蛋白A結(jié)合使IAA抗體修飾到芯片上,最后進行對IAA的檢測。傳感器2:先在裸金片上修飾HDT,Au/Ag合金納米粒子可與金硫鍵、金銀鍵結(jié)合從而使其修飾到芯片上,接下來在傳感器2上依次修飾MPA、蛋白A、IAA抗體的方法與傳感器1相同。結(jié)果兩種傳感器均能成功的檢測IAA,傳感器1對IAA濃度為175~350μg/L所對應(yīng)的波數(shù)變化進行線性回歸分析,得到線性方程為y=-6.58+0.103 x,R2=0.994,檢測限為25μg/L(S/N=3);傳感器2對IAA濃度為17.5~250μg/L所對應(yīng)的波數(shù)變化進行線性回歸分析,得到線性方程為y=4.27+0.104 x,R2=0.999,檢測限為2.2μg/L(S/N=3)。說明傳感器2中引入Au/Ag合金納米粒子確實可以有效的提高檢測的靈敏度,此外本實驗所構(gòu)建的SPR生物傳感器具有良好的穩(wěn)定性,特異性及重現(xiàn)性。(2)構(gòu)建兩種SPR生物傳感器并成功檢測ABA:一種是MPA構(gòu)建的SPR生物傳感器(傳感器1),另一種是巰基己酸構(gòu)建的SPR生物傳感器(傳感器2)。通過分子自組裝技術(shù)對SPR生物傳感器的芯片進行修飾。傳感器1依次修飾HDT、Au/Ag合金納米粒子、MPA、蛋白A、IAA抗體,最后對ABA進行檢測。傳感器2與傳感器1的不同之處在于將傳感器1的MPA換成了巰基己酸,增長了3個碳鏈,通過加長檢測鏈的方法來提高SPR生物傳感器的靈敏度。結(jié)果兩種SPR生物傳感器均能成功的檢測ABA,傳感器1對ABA濃度為13.1~23.8μg/L所對應(yīng)的波數(shù)變化進行線性回歸分析,得線性方程為y=1.29+0.074 x,R2=0.995,檢測限為2.3μg/L(S/N=3);傳感器2對ABA濃度為3.3~40μg/L所對應(yīng)的波數(shù)變化進行線性回歸分析,得線性方程為y=1.12+0.044 x,R2=0.992,檢測限為0.56μg/L(S/N=3)。說明傳感器2通過加長檢測鏈確實可以有效提高檢測的靈敏度,此外本實驗所構(gòu)建的SPR生物傳感器具有良好的精密度,穩(wěn)定性,特異性及重現(xiàn)性。
[Abstract]:3- indoloacetic acid (IAA) and abscisic acid (ABA) are common plant hormones and play an important role in plant growth and development. The basic function of.IAA not only accelerates and regulates the growth and development of plants, but also hinders plant growth and root, stem and group formation, the basic role of.ABA is to promote and inhibit planting. At present, most of these two kinds of plant hormone detection methods are complicated, time-consuming, expensive, high cost, and poor selectivity. Therefore, this study uses the advantages of real-time monitoring, samples without preprocessing, rapid and sensitive detection and so on. In order to improve the sensitivity of IAA and ABA, two kinds of plant hormones are detected by the SPR biosensor. In order to improve the sensitivity of SPR sensors, this paper adopts two methods: one is to introduce gold / silver alloy nanoparticles into SPR biosensors as sensitized materials. The other is to improve the sensitivity of SPR biosensor detection by the way of lengthening the detection chain. This article includes two parts of the experiment: (1) the construction of two kinds of SPR biosensors and the detection of IAA: one is the traditional mode SPR biosensor (sensor 1), the other is the SPR biosensor (sensor 2) which introduces the Au/Ag alloy nanoparticles as the sensitizing material (the sensor 2). First, the SPR chip is modified with different substances. 1: first modified MPA on bare gold slices and then modified protein A. Protein A can be combined with the carboxyl of EDC/NHS at the end of MPA to modify the protein A on the chip and inactivate the chip with ethanolamine hydrochloride. The non antigen binding site of IAA antibody can be combined with protein A to modify the IAA antibody to the chip. Finally, IAA is detected. Sensor 2:. First, the HDT is modified on the bare gold plate. The Au/Ag alloy nanoparticles can be combined with gold and silver bonds with gold and silver bonds to make them modified to the chip. Then, the MPA, protein A, IAA antibody are the same as the sensor 1. The results of the two sensors are all successful in detecting IAA, and sensor 1 to the IAA concentration corresponding to 175~350 u g/L. Linear regression analysis is carried out, the linear equation is y=-6.58+0.103 x, R2=0.994, the detection limit is 25 g/L (S/N=3); sensor 2 is linear regression analysis on the wave number of IAA concentration corresponding to 17.5~250 u g/L, and the linear equation is y=4.27+0.104 x, R2=0.999, and the detection limit is 2.2 micron g/L. The alloy nanoparticles can effectively improve the sensitivity of the detection. In addition, the SPR biosensor constructed by this experiment has good stability, specificity and reproducibility. (2) two kinds of SPR biosensors are constructed and ABA: is a SPR biosensor (sensor 1) constructed by MPA, and the other is the SPR generation of mercapto hexanoacid. The sensor (sensor 2). Modifies the SPR biosensor chip by molecular self-assembly technology. Sensor 1 modifies HDT, Au/Ag alloy nanoparticles, MPA, protein A, IAA antibody in turn, and finally detects ABA. The difference between sensor 2 and sensor 1 is that the MPA of the sensor 1 is replaced by mercapto hexanoprol acid and 3 carbon chains are increased. The sensitivity of the SPR biosensor was enhanced by the method of lengthening the detection chain. Results two kinds of SPR biosensors were able to detect ABA successfully. Sensor 1 had linear regression analysis on the wave number changes corresponding to the ABA concentration of 13.1~23.8 mu g/L. The linear equation was y=1.29+0.074 x, R2= 0.995, and the detection limit was 2.3 mu g/L (S/N=3); sensor 2 pairs of ABA. The linear regression analysis of the wave number variation corresponding to 3.3~40 mu g/L is obtained. The linear equation is y=1.12+0.044 x, R2=0.992, and the detection limit is 0.56 mu g/L (S/N=3). It shows that the sensor 2 can effectively improve the sensitivity of the detection by adding a long detection chain. Furthermore, the SPR biosensor built in this experiment has good precision and stability. Specificity and reproducibility.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TP212.3

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本文編號：2015125