Dissertation > Excellent graduate degree dissertation topics show

Preparation of Monoclonal Antibody Against PR-39 of Piglets and Research on Regulation of It’s Gene Expression

Author: AnZhongHua
Tutor: WangYiZhen
School: Zhejiang University
Course: Animal Nutrition and Feed Science
Keywords: Weaned piglets PR-39 Lactoferrin Hybridoma technology Monoclonal antibody Enzyme-linked immunosorbent assay Gene Expression
CLC: S852.4
Type: Master's thesis
Year: 2007
Downloads: 18
Quote: 0
Read: Download Dissertation

Abstract


In this study, the synthesized partial amino acid of PR-39, that contains it’s reactive sites was used to establish the monoclonal antibody (McAb) against PR-39 by hybridoma technology. Then the monoclonal antibody was used to develop the effect of lactoferrin on PR-39 gene expression at translation level by inhibition enzyme-linked assay (ELISA).1.The synthesized partial amino acid of PR-39 FFPPRLPPRIP (PR11),containing it’s reactive sites, was conjugated to ovalbumin (OVA) and bovine serum albumin (BSA) using the coupling procedure in order to prepare artifical immunogen PR11-OVA and coating antigen PR11-BSA by carbodiimide method. Then the hybridoma cell strains named 3H5 and 5H7 that secreted monoclonal antibody (McAb) against antibacterial peptide PR-39 were established using hybridoma technology. These hybridoma cell strains were injected intraperitoneally into Balb/c mice respectively to produce ascites of monoclonal antibody. Each ascites was purified by caprylic acid and ammonium sulfate.2.The obtained McAb was evaluated the immunological properties.The results showed that the concentrations of IgG in these ascites were 28.463mg/ml and 16.124mg/ml and the indirect enzyme-linked immunosorbent assay(ELISA) titer were both higher than 106. The isotypes of the McAb were IgG1 by double immunodiffusion test.3.The bone marrow cells which was separated from the femur of 30 days weaned piglets (Duroc×Landrace×Yorkshine) were sampled for 48h, then induced by lactoferrin (0,10,100, 1000mg/ml) for 3h,6h,12h,24h, respectively, in order to obtain cell culture supernatants containing antibacterial peptide PR-39.The concentrations of PR-39 in these culture supernatants were measured to determine the effect of lactoferrin on PR-39 gene expression at translation level by inhibitive enzyme-linked immunosorbent assay using the McAb against PR-39 with high activity and specificity. The results showed that the concentration of PR-39 in the culture supernatants of bone marrow cells of piglets induced by lactoferrin for 3h and 6h increased with the increasing concentration of lactoferrin and the concentration of PR-39 increased significantly (P<0.05)when the concentration of lactoferrin was 1000mg/ml, suggesting that lactoferrin may enhance PR-39 gene expression at translation level.However the expression of PR-39 had no significant differences between the trials and the control groups at 12h or 24h and the concrete mechanism needs further study.In this study the bone marrow cells were induced by lactoferrin under different concentrations to study the effect of lactoferrin on PR-39 gene expression at translation level.The results indicated that lactoferrin could enhance PR-39 gene expression when the cells were induced for 3h or 6h, which were in agreement with the results at transcription level.These results suggested that antibacterial peptide gene expression of weaned piglets may be enhanced by adding some nutriments or physical active substances in feedstuff, such as lactoferrin, to improve the piglet’s immunity. This is very important for weaned piglet to enhance the resistance to the environmental pathogenic bacterias. Furthermore, affinity chromatography method, a new convenient method may be established to purify antibacterial peptide PR-39 by these obtained monoclonal antibodies. The method and thought in this study layed the theoretical foundation for the research on other small molecule peptide as well.

Related Dissertations

  1. Research and Implementation on Knowledge Representation of Gene Regulatory Network,Q78
  2. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  3. The Research of the High-Rise Building’s Form and Expression in the New Century,TU971
  4. Research on Design of Teaching Buildings for Arts Departments of Higher Learning Institutions,TU244.3
  5. Research on Temporal Information Recognition and Normalization,TP391.1
  6. Expression of D-AtCGS in E. Coli and Preparation of Polyclonal Antibody Against D-AtCGS,Q943.2
  7. Cloning and Expression of CHS and CHI Genes and Their Regulation on the Accumulation of Flavonoids in ’Cara Cara’ Navel Orange (Citrus Sinensis Osbeck) and ’Guoqing NO.4’ Satsuma Mandarin (Citrus Unshiu Marcow),S666.4
  8. The Study on Arabidopsis Thaliana Heat Shock Factor HSFA 1d Response to Formaldehyde Stress,Q945.78
  9. Expression of hBMP4 and hBMP7 in Chinese Hamster Ovary Cells,Q78
  10. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  11. Study on Bispecific Monoclonal Antibody to Pesticide Imidacloprid and Parathion-Methyl,S482.2
  12. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  13. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  14. Chemical Structures and Mechanism Research of Anti-aging Leading Compounds from Natural Products,R285.5
  15. Expression of Prolyl Isomerase Pin1 in Osteosarcoma and the Effect of Regulation on Cell Cycle,R738.1
  16. CYP2C19 Gene Polymorphism in Patients with Ischemic Cerebrovascular,R743
  17. The Expression Survey of BMP Signalling Pathway in the Human Embryonic Tooth Germ,R78
  18. Preliminary Study of Phosphorylation Effect on UGT1A3 Metabolism Activity Towards Drugs,R346
  19. Cloning and Expression of Growth Hormone Gene in Cobia (Rachycentron Canadus),Q786
  20. Expression and Purification of Influenza a Virus M2 Protein and Detection of Its Antigenicity,R392
  21. Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap,S917.4

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Immunology
© 2012 www.DissertationTopic.Net  Mobile