Syntheses of immunodominant peptide regions of hepatitis C viral pathogens using PS-BDODMA resin: a single peptide derived from the conserved domain (E2/NS1) was highly effective in detecting anti-HCV antibodies.
Peptides were synthesized with very high purity and yield on a highly solvating copolymer of 1,4-butanediol dimethacrylate cross-linked polystyrene support (PS-BDODMA). The polymer was synthesized by radical aqueous suspension polymerization using toluene as the diluent and 1% polyvinyl alcohol as the suspension stabilizer. The supports were highly resistant to various chemical reagents and solvents that are frequently used in polypeptide synthesis. The peptides synthesized on this support were designed from the core (C), envelope (E1 and E2) and nonstructural protein (NS1-NS5) regions of the prototype hepatitis C viral (HCV) polyprotein, and were used to develop a peptide-based immunoassay (PBEIA) for the detection of HCV antibodies. The purity of these peptides was tested by HPLC. Peptide identity was confirmed by amino acid analysis and MALDI-TOF-MS. A single peptide chosen from a conserved area (E2/NS1) at the C-terminus of the hypervariable region (HVRI) was found to be sufficient for effective and reliable diagnosis of HCV infection in infected individuals, as well as also apparently healthy individuals. The CD spectrum of the peptide shows no preference for any ordered secondary structure. When used, peptide mixtures from various protein regions of HCV reduced the sensitivity and reliability of the diagnosis partly because of epitope masking.
Kumar
K S
KS
Rajiv Gandhi Centre for Biotechnology, Jagathy, Thiruvananthapurum, India.
Roice
M
M
Sasikumar
P G
PG
Poduri
C D
CD
Sugunan
V S
VS
Rajasekharan Pillai
V N
VN
Das
M R
MR
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D016174:Hepacivirus / Q000737:chemistry*; D018937:Hepatitis C Antibodies / Q000032:analysis* / Q000276:immunology; D016056:Immunodominant Epitopes / Q000737:chemistry* / Q000276:immunology; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D011137:Polystyrenes / Q000737:chemistry*
D018937:Hepatitis C Antibodies; D016056:Immunodominant Epitopes; D011137:Polystyrenes