Advantages of branched peptides in serodiagnosis. Detection of HIV-specific antibodies and the use of glycine spacers to increase sensitivity.
The reactivities of antibodies with branched and monomeric peptides were compared in ELISA assays. We found that lower amounts of antibodies could be detected with branched peptides than with monomeric peptides. This was observed with a monoclonal antibody and with antibodies in the sera of various HIV-positive individuals. To investigate the physical aspects of branched peptides important for the observed increase in sensitivity, glycine spacers of different lengths were introduced between the branched lysine core and the epitope reacting with the monoclonal antibody. The effect of the number of glycine residues, both on the sensitivity of antibody detection and on the amount of branched peptide needed to produce a given signal, was studied and the optimum was found at 4-5 residues. We discuss the basis for these findings and conclude that the routine use of branched peptides for serodiagnosis will give both greater sensitivity and appreciable cost savings.
Marsden
H S
HS
MRC Virology Unit, Institute of Virology, Glasgow, UK.
Owsianka
A M
AM
Graham
S
S
McLean
G W
GW
Robertson
C A
CA
Subak-Sharpe
J H
JH
D015492:AIDS Serodiagnosis; D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis*; D000595:Amino Acid Sequence; D000818:Animals; D005998:Glycine; D015483:HIV Antibodies / Q000032:analysis*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D011817:Rabbits
D015483:HIV Antibodies; D010446:Peptide Fragments; D005998:Glycine