Antibodies against the two major surface glycoproteins of bovine respiratory syncytial virus (BRSV), G and F, play a role in protection against BRSV-associated disease, but only the antibody response against the F protein has been well described. Therefore, we used a novel peptide-based enzyme-linked immunosorbent assay (G peptide-ELISA) to compare immunoglobulin G (IgG) and IgG subclass antibody responses against the G protein with the antibody response against the F protein, as measured by a conventional BRSV ELISA (F-ELISA). Experimental infection of cattle induced significantly lower antibody titers than did natural infection. After natural primary infection, G peptide-specific antibodies declined more rapidly and to lower levels than the F protein-specific antibodies. As a consequence, the G peptide-ELISA detected more reinfections than did the F-ELISA. Ratios of G- and F-specific IgG1/IgG2 antibody titers did not differ markedly after infection or vaccination. Interestingly, after natural infection calves did not develop an IgG2 response to the complete G protein. In contrast, adult cattle had high IgG2 titers against this protein. Vaccination with a live vaccine induced low antibody titers, similar to the titers after experimental infection, whereas vaccination with an inactivated vaccine induced high titers. The results indicate that the kinetics of the G- and F-specific antibody responses differ. Furthermore, the IgG subclass response against the unglycosylated central region of the G protein is similar to the IgG subclass response to the F protein, but the IgG subclass response differs from the response to the complete G protein.