Assessment of Surfactant Protein A (SP-A) dependent agglutination

BMC Pulmonary Medicine

Table 3 Tests for SP-A self-agglutination-assay dependency

Beads	Goat-anti-human-SP-A antibody	Anti-goat antibody	Serum	Ca²⁺	Agglutination
+	+	+	+	-	-
+	+	+	+	+	+
+	+	+	-	+	-
+	+	+	-	-	-
+	-	+	+	+	-
+	-	-	+	+	-
-	+	+	+	+	-
+	+	+	SP-D	-	-
+	+	+	SP-D	+	-
+	+	+	-	+ with Mannose	-
+	+	+	+	+ with Mannose	-
+	+	+	+/without SP-D	+	+
+	+	+	+/+Triton X	+	-

The table shows twelve conditions of the bead assay with varying reagents: beads, anti-goat antibody, goat-anti-human antibody, human serum and calcium ions (Ca²⁺). The "+" means that the reagent was used for incubation. Without SP-A and/or calcium ions in serum, and also with Mannose no agglutination was detected. The experiments were repeated three times. For the last condition a concentration of 0.1% (v/v) Triton X-100 was used (0.3 μl Triton X-100 in 1 ml pure water, 3 μl of this water were used for the assay). A final concentration of 100 ng/ml SP-A (1 ng) in test serum and 100 ng/ml SP-D standard (1 ng recombinant human SP-D) were used.

ISSN: 1471-2466