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Preparation and Detection of Chicken Egg Yolk IgY Antibody Against Non-small Cell Lung Cancer (NCLC) Correlated Peptide by ELISA

Author: YangJian
Tutor: WangWenJing；WangWeiRong；LuWei
School: Donghua University
Course: Biochemistry and Molecular Biology
Keywords: IgY technology synthesis peptide antibody against peptide detection by ELISA lung cancer diagnosis
CLC: R734.2
Type: Master's thesis
Year: 2010
Downloads: 38
Quote: 1
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Abstract


Lung cancer is a common malignant tumor,80%of which is non-small cell lung cancer (NSCLC),with the increasing prevalence of morbility and mortality in the world.It is very difficult to do an early diagnosis as NSCLC.It is to late to gain a operation chance to cure lung caner when 80%patients diagnosed with a advanced stage.And to do a radiation therapy or a chemotherapy offers little help.Survival Rate in 5 years is no more than 15%.On the othe hand there are 70%of Survival Rate in 5 years after an operation on early stage,with the early diagnosis rate only about 15%.We obtained two peptides with high sensitivity and specificity of NSCLC for early diagnosis when use a surface-enhanced laser desorption/ionization time-of-flight mass spectrometry.And we do the research this time to develop more than before.Peptide is a kind of a chemical compounds,and the molecular structural of it have been detected between amino acid and protein.Also peptide is the structure and function segment of the protein.It is made up of amino acids.Like mammals,chickens protect their offspring by transferring maternal antibodies their offspring by transferring maternal antibodies from serum to egg ylok.The properties of the 7S Ig species from avians are slightly different from those of mammalian IgG,and are called IgY.The protein has an Mr of 180,000,which consists of two subunits:a heavy chain of 67,000-70,000 and a light chain of 22,000-30,000.The advantage is that conserved mammalian proteins are usually more immunogenic in the phylogenetically distant birds.[Objective]To prepare chicken egg yolk IgY antibody against non-small cell lung cancer (NSCLC)-related peptide and to detect lung cancer by ELISA with specific IgY antibody.[Methods] Yolk immunoglobulin(IgY) was obtained by injecting NSCLC-related peptide(the length of two peptide are 36AA and 64AA) bound to KLH into the hens.A relatively high purity and specific activity of IgY was obtained by water dilution-caprylie acid and ammonium sulfate precipitation and affinity chromatograph.An optimal working condition is established by the way of chessboard titration.The titres of specific IgY antibody were determined by indirect ELISA.Blood sera samples including 96 lung cancer and 102 normal were examined by the combination of two specific IgY antibody.[Results]The titers of the two peptide were 1:104 and 1:105 by ELISA.A sensitivity of 60.4%and a specificity of 81.4%are obtained after detecting the sera samples including 96 lung cancer and 102 normal.[Conclusion]Diagnosing lung cancer by IgY technology may have a good clinical value.

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CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
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