Selection of Specific Peptides for Coccidioides spp. Obtained from Antigenic Fractions through SDS-PAGE and Western Blot Methods by the Recognition of Sera from Patients with Coccidioidomycosis

Molecules. 2018 Nov 30;23(12):3145. doi: 10.3390/molecules23123145.

Abstract

Antigenic fractions of 100, 50, 37, and 28 kDa obtained through the SDS-PAGE method that were more frequently recognized by anti-Coccidioides antibodies in the sera of coccidioidomycosis patients were selected using western blotting. Subsequently, these bands were sequenced, and the obtained proteins were analysed by BLAST to choose peptides specific for Coccidioides spp. from among the shared aligned sequences of related fungi. A peptide specific for C. immitis was selected from the "GPI anchored serine-threonine rich protein OS C. immitis", while from the "uncharacterized protein of C. immitis", we selected a peptide for C. immitis and C. posadasii. These proteins arose from the 100 kDa antigenic fraction. From the protein "fatty acid amide hydrolase 1 of C. posadasii" that was identified from the 50 kDa antigenic fraction, a peptide was selected that recognized C. immitis and C. posadasii. In addition, the analysis of all the peptides (353) of each of the assembled proteins showed that only 35 had 100% identity with proteins of C. immitis and C. posadasii, one had 100% identity with only C. immitis, and one had 100% identity with only C. posadasii. These peptides can be used as diagnostic reagents, vaccines, and antifungals.

Keywords: Coccidioides spp.; SDS-PAGE; coccidioidomycosis; immunodiagnosis; peptides.

MeSH terms

  • Adult
  • Aged
  • Amino Acid Sequence
  • Antigens, Fungal / chemistry
  • Antigens, Fungal / isolation & purification*
  • Blotting, Western / methods*
  • Child
  • Coccidioides / immunology*
  • Coccidioides / isolation & purification
  • Coccidioidomycosis / blood*
  • Coccidioidomycosis / immunology*
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Peptides / chemistry
  • Peptides / isolation & purification*
  • Young Adult

Substances

  • Antigens, Fungal
  • Peptides