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Development and evaluation of a flow...

Ho, Ja-an Annie.

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Development and evaluation of a flow injection liposome immunoanalysis (FILIA) system for fumonisin B1.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Development and evaluation of a flow injection liposome immunoanalysis (FILIA) system for fumonisin B1./
作者:	Ho, Ja-an Annie.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 1998,
面頁冊數:	189 p.
附註:	Source: Dissertations Abstracts International, Volume: 60-04, Section: B.
Contained By:	Dissertations Abstracts International60-04B.
標題:	Analytical chemistry. -
電子資源:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9838787
ISBN:	9780591923902

Development and evaluation of a flow injection liposome immunoanalysis (FILIA) system for fumonisin B1.
Ho, Ja-an Annie.

Development and evaluation of a flow injection liposome immunoanalysis (FILIA) system for fumonisin B1. - Ann Arbor : ProQuest Dissertations & Theses, 1998 - 189 p.

Source: Dissertations Abstracts International, Volume: 60-04, Section: B.

Thesis (Ph.D.)--Cornell University, 1998.

This item must not be sold to any third party vendors.

Fumonisins are a group of mycotoxins, discovered in 1988 from cultures of Fusarium moniliforme strain MRC 826, which are known to cause leukoencephalomalacia (LEM) in horses, pulmonary edema syndrome (PES) in swine, and to be highly toxic to a variety of experimental animals. Epidemiological evidence also shows there is a possible correlation between human esophageal cancer and fumonisins. Additionally, fumonisins are structurally similar to sphingosine and may exert their biological activity through their ability to block sphinganine- and sphingosine-N-acyltransferases, which are involved in sphingolipid biosynthesis. The objective of this research was to develop a FILIA system, as an alternative to current analytical methods, that would provide higher sensitivity, increased speed, lower cost, and improved simplicity for the quantitation of fumonisin B1 (FmB1) in food products and animal feeds. Liposomes, which encapsulate sulforhodamine B as signal amplifiers in FILIA, provide instantaneous enhancement rather than time-dependent enhancement as in enzyme immunoassays (such as ELISA). In this research, a method was developed for the preparation of FmB1-tagged liposomes by coupling maleimide-derivatized FmB1 to the sulfhydryl groups incorporated in the bilayer of the liposomes. The synthesis methods for FmB1-KLH (keyhole limpet hemocyanin) conjugates used as immunogens to raise polyclonal antibodies in Rambouilet sheep were also studied. The immunoassay was based on competitive binding of FmB1 and FmB1-tagged liposomes to the limited number of immobilized anti-FmB1 antibody binding sites in a capillary immunoreactor column (50 cm x 0.53 mm i.d.). The fluorescent signal generated after lysis of the antibody-bound liposomes is inversely proportional to the concentration of free FmB1 present in the sample. Recovery studies indicated that a rate of 80-92% could be obtained when commercial yellow cornmeal spiked with FmB1 was extracted in 75% methanol, which compared favorably with the result of 86-90% obtained by using high-performance liquid chromatography (HPLC). An assay could be performed in $<$11 minutes, and filtration was the only step needed for sample preparation. The calibration curve for FmB1 in Tris-buffered saline solution had a working range of 0.1-100 ng, and the limit of detection of 0.1 ng was also found. This work demonstrates the feasibility of using flow injection liposome immunoanalysis for the detection of fumonisin B1, which offers the advantages of high specificity, sensitivity, speed, and simplicity of operation.

ISBN: 9780591923902Subjects--Topical Terms:

3168300
Analytical chemistry.
Subjects--Index Terms:

Flow injection liposome immunoanalysis

Development and evaluation of a flow injection liposome immunoanalysis (FILIA) system for fumonisin B1.
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520 $a Fumonisins are a group of mycotoxins, discovered in 1988 from cultures of Fusarium moniliforme strain MRC 826, which are known to cause leukoencephalomalacia (LEM) in horses, pulmonary edema syndrome (PES) in swine, and to be highly toxic to a variety of experimental animals. Epidemiological evidence also shows there is a possible correlation between human esophageal cancer and fumonisins. Additionally, fumonisins are structurally similar to sphingosine and may exert their biological activity through their ability to block sphinganine- and sphingosine-N-acyltransferases, which are involved in sphingolipid biosynthesis. The objective of this research was to develop a FILIA system, as an alternative to current analytical methods, that would provide higher sensitivity, increased speed, lower cost, and improved simplicity for the quantitation of fumonisin B1 (FmB1) in food products and animal feeds. Liposomes, which encapsulate sulforhodamine B as signal amplifiers in FILIA, provide instantaneous enhancement rather than time-dependent enhancement as in enzyme immunoassays (such as ELISA). In this research, a method was developed for the preparation of FmB1-tagged liposomes by coupling maleimide-derivatized FmB1 to the sulfhydryl groups incorporated in the bilayer of the liposomes. The synthesis methods for FmB1-KLH (keyhole limpet hemocyanin) conjugates used as immunogens to raise polyclonal antibodies in Rambouilet sheep were also studied. The immunoassay was based on competitive binding of FmB1 and FmB1-tagged liposomes to the limited number of immobilized anti-FmB1 antibody binding sites in a capillary immunoreactor column (50 cm x 0.53 mm i.d.). The fluorescent signal generated after lysis of the antibody-bound liposomes is inversely proportional to the concentration of free FmB1 present in the sample. Recovery studies indicated that a rate of 80-92% could be obtained when commercial yellow cornmeal spiked with FmB1 was extracted in 75% methanol, which compared favorably with the result of 86-90% obtained by using high-performance liquid chromatography (HPLC). An assay could be performed in $<$11 minutes, and filtration was the only step needed for sample preparation. The calibration curve for FmB1 in Tris-buffered saline solution had a working range of 0.1-100 ng, and the limit of detection of 0.1 ng was also found. This work demonstrates the feasibility of using flow injection liposome immunoanalysis for the detection of fumonisin B1, which offers the advantages of high specificity, sensitivity, speed, and simplicity of operation.
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