FIG. 4.

Analysis of binding of gD to HveCt by ELISA. Ninety-six-well plates were saturated with 50 μl of 200 nM HveCt in PBS and incubated with variable concentrations of purified HSV glycoproteins. (A) Glycoproteins bound to immobilized HveC were detected with specific antibodies (R7 for gD, R47 for gC, R69 for gB, and R137 for gH-gL) followed by peroxidase-conjugated secondary antibody and substrate. (B) gD-1(306t)KOS and gD-2(306t)333 at various concentrations were incubated on HveCt-coated plates. (C) Mutant gD (QAAt) lacking N-CHO was compared to the glycosylated control gD(306t) for binding to immobilized HveCt. (D) Purified gD-1(306t) was reduced and alkylated prior to incubation on the HveCt-coated plate. Rabbit polyclonal serum R7 was used to detect any type of gD.