The influence of pH and ionic strength on the coating of peptides of herpes simplex virus type 1 in an enzyme-linked immunosorbent assay.
Rabbits were immunized with synthetic peptides of herpes simplex virus type 1 glycoproteins, coupled to a carrier protein with glutaraldehyde. Antibodies directed against the peptides were determined in an enzyme-linked immunosorbent assay (ELISA). Either free peptides or peptides coupled with glutaraldehyde to another carrier protein than the one used for immunization were used as the coating antigen. When conjugated peptides were used as the coat, it was necessary in some instances to correct the antibody titers for a substantial amount of antibody activity against glutaraldehyde. When free peptides were used, optimal coating conditions with regard to pH and ionic strength had to be determined, since some peptides failed to coat under standard conditions, at pH 9.6. The results show that some peptides needed stringent pH conditions while others could be coated in a broad pH range. The addition of 0.6 M NaCl had a favorable effect on peptide coating.
Geerligs
H J
HJ
Laboratorium voor Medische Microbiologie, Rijksuniversiteit Groningen, The Netherlands.
Weijer
W J
WJ
Bloemhoff
W
W
Welling
G W
GW
Welling-Wester
S
S
D000937:Antigen-Antibody Reactions; D000956:Antigens, Viral / Q000032:analysis*; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010455:Peptides / Q000276:immunology*; D018139:Simplexvirus / Q000276:immunology*; D012965:Sodium Chloride / Q000494:pharmacology
D000956:Antigens, Viral; D010455:Peptides; D012965:Sodium Chloride