Intracellular polysaccharide fractions were isolated from calcifying B‐type cells of Emiliania huxleyi and separated by electrophoretic fractionation. In all fractions, the polysaccharide was immunologically related to the polysaccharide of (extracellular) B‐type coccoliths (CP‐B) and not to polysaccharides of A‐type coccoliths (CP‐A). Most polysaccharide fractions also contained protein material. The fraction with the largest proportion of protein was used to raise antibodies. The resulting antiserum, α‐BP, contained antibodies against both CP‐B‐ and protein‐epitopes. The antibodies specific for polysaccharide‐epitopes reacted with intracellular polysaccharide fractions of B‐type cells only. In contrast, the antibodies specific for protein‐epitopes reacted with the intracellular fractions of B‐type as well as A‐type cells. With immunolocalization, the presence of protein antigen in a layer surrounding both types of cells was demonstrated. A cDNA library of E. huxleyi was screened with α‐BP, and a gene called gpa was isolated. The open reading frame of gpa was found to encode a protein (GPA) of 36,608 D, containing, inter alia, 24% acidic residues (18% glutamic acid and 6% aspartic acid), 12% proline, and 23% alanine. GPA has two repeats, one containing a sequence resembling the Ca2+‐binding loop of EF‐hands. Overproduction of GPA in a prokaryotic system yielded a dimeric product capable of binding Ca2+. The possible role of GPA in the formation of coccoliths in E. huxleyi is discussed.

Production of coccoliths by cells of Emiliania huxleyi (Lohmann) Hay and Mohler was measured during exposure of the cells to two diel light‐dark cycles (16:8 h). During the light period about eight coccoliths per cell were formed at a constant rate of one coccolith per 2 h. Cells divided during the first half of the dark period. No coccolith production took place during the dark period. With electron microscopy we found early‐stage, coccolith‐production compartments in cells after mitosis while still in the dark. No calcification was observed in these compartments. Cells grown on enriched seawater (Eppley's medium) tended to produce enough coccoliths to cover the cell in a single layer. When these cells reached the stationary phase coccolith production stopped. Coccolith production was induced by removal of extracellular coccoliths. Cells grown on medium containing 2% of the nitrate and phosphate of Eppley's medium tended to produce coccoliths in the stationary phase. This resulted in the formation of multiple layers of coccoliths. The multiple covering was restored after decalcification of stationary cells. Formation of multiple layers of coccoliths may help the cells reach deeper, nutrient‐rich water by increasing the sinking rate of the cells.

Immunoglobulin M antibodies against a major serological antigen (Mr 6 kD) were detected in human sera by indirect ELISA, antibody capture ELISA, and immunoblot test. In contrast to indirect ELISA, the immunoblot test gave no false positive reactions, not even with those sera containing a high level of rheumatoid factor. However, the immunoblot test gave false negative results with sera which gave positive results in both ELISA tests. The antibody capture ELISA gave no false negative reactions. All positive sera except one reacted specifically with Toxoplasma gondii antigens.