Table 1.

Antibody titers by Enzyme-Linked ImmunoSorbent Assay (ELISA)^a

Mouse #	Globo-H Ceramide		GM2 Ceramide		OSM for sTn		dPSM for TF		dOSM for Tn
	IgG	IgM	IgG	IgM	IgG	IgM	IgG	IgM	IgG	IgM
Pre-Serum	0	0	0	0	0	0	0	160	0	0
1	160	160	160	0	400	320	400	400	800	0
2	640	160	640	0	1600	40	400	400	1600	0
3	640	160	640	0	800	320	200	100	6400	200
4	640	640	2560	0	1600	2560	1600	800	3200	400
5	640	160	640	0	1600	20	1600	800	1600	400
Median	640	160	640	0	1600	320	800	400	1600	200

^aIgM or IgG antibody reciprocal titers of pre- and post-vaccination sera tested for the five antigens in the pentavalent vaccines. ELISA assays were performed to determine IgM and IgG serum antibody titers as previously described.⁹ In brief, Globo-H ceramide, GM2 ceramide, ovine submaxillary mucin (OSM expressing sTn), desialylated ovine submaxillary mucin (dOSM expressing Tn), or desialylated porcine submaxillary mucin (dPSM expressing TF) was coated on ELISA plates at an antigen dose of 0.1 μg/well and incubated overnight at 4 °C. Nonspecific sites were blocked with 1% human serum albumin (HSA) for 2 h, and serially diluted antiserum was added to each well. After 1 h of incubation, the plates were washed, and alkaline phosphatase labeled goat anti-mouse IgM or IgG was added at 1:400 dilution (Southern Biotechnology Associates Inc., Birmingham, AL). The antibody titer was defined as the highest dilution with absorbance of 0.1 or greater over that of normal control mouse sera.