Standing out in the field of IgY Immunotechnology

  • Home
    Home A full collection of all the Research Archive entries.
  • Years
    Years Sort entries by year.
  • Tags
    Tags Displays a list of tags that have been used in the blog.
  • Archives
    Archives Contains a list of research entries that were created previously.

Evaluation of recombinant Cryptosporidium hominis GP60 protein and anti-GP60 chicken polyclonal IgY for research and diagnostic purposes.

Posted by on in 2017
  • Font size: Larger Smaller
  • Hits: 261
  • Print
Miura VC1,2Aoki SM2Peitl P Junior2Pires LC2Dalmagro P2Nakamura AA1Meireles MV1. 2017.  Rev Bras Parasitol Vet.  26(2):205-210. doi: 10.1590/S1984-29612017032. Epub 2017 Jun 22.

Abstract

In this study, a method for expressing Cryptosporidium hominis GP60 glycoprotein in Escherichia coli for production of polyclonal anti-GP60 IgY in chickens was developed aiming future studies concerning the diagnosis, prevention and treatment of cryptosporidiosis. The full-length nucleotide sequence of the C. hominis gp60 gene was codon-optimized for expression in E. coli and was synthesized in pET28-a vector. Subcloning was performed on several different strains of BL21 E. coli. Temperature, time and inducer IPTG concentration assays were also performed and analyzed using SDS-PAGE. The optimal conditions were observed at a temperature of 37 °C, with overnight incubation and 1 mM of IPTG. Purification was performed by means of affinity chromatography using the AKTA Pure chromatography system and the Hi-Trap™ HP column (GE Healthcare). The recombinant protein GP60 (rGP60) thus generated was used to immunize laying hens owing the production of polyclonal IgY. Western blot and indirect immunofluorescence showed that the polyclonal antibody was capable of binding to rGP60 and to Cryptosporidium parvum sporozoites, respectively. The rGP60 and the IgY anti-rGP60 generated in this study may be used as templates for research and for the development of diagnostic methods for cryptosporidiosis.

PMID:
 
28658417
 
DOI:
 
10.1590/S1984-29612017032
Free full text
Last modified on