Purification and binding properties of porcine plasma ficolin that binds Actinobacillus pleuropneumoniae

Cloning, binding properties, and tissue localization of rainbow trout (Oncorhynchus mykiss) ladderlectin

“…Direct plasma binding assays [41,42] were used to determine the ability of RTLL to bind to relevant targets such as purified crab chitin, Artemia cysts and intact bacteria. Under the serum incubation and wash conditions employed to detect lectins that could bind to intact organisms, various bands including immunoglobulins and pentraxins elute with saccharides [35].…”

“…Ligand-coupled matrices were washed with 500 ml of milliQ (deionized and 0.22 mm filtered) water and equilibrated in TBSeCa þþ . The production of an N-acetyl Toyopearl AF-Amino 650M matrix was prepared as previously described [40,41]. Briefly, 10 mg of Toyopearl AF-Amino 650M was washed with 1 l of distilled water and incubated for 30 min at 4 C following the addition of 4 ml of acetic anhydride and 8 ml of 0.2 M sodium acetate.…”

“…salmonicida Protocols developed for the isolation of rainbow trout plasma proteins that bound to purified crab chitin, Artemia cysts and intact A. salmonicida subsp. salmonicida (ASAL) bacteria were based upon those previously described [41,42]. Chitin is a long-chain polysaccharide constructed of repeating units of N-acetylglucosamine and is the major component of the exoskeletons of parasitic arthropods such as sea lice Lepeophtheirus salmonis.…”

Bacterial-binding activity and plasma concentration of ladderlectin in rainbow trout (Oncorhynchus mykiss)

“…Bacterial-binding assays [30,31] were performed using clinical isolates of A. hydrophila, Pseudomonas sp., two A. salmonicida subsp. salmonicida isolates and Yersinia ruckeri (ATCC 29473).…”

Expression of recombinant Atlantic salmon serum C-type lectin in Drosophila melanogaster Schneider 2 cells

“…salmonicida strain A449 isolated from brown trout (Michel 1979) was obtained from the National Research Council of Canada (Halifax, NS). The bacteria were cultured in tryptic soy broth at 16°C for 48-72 h. Recombinant lectin binding to A. salmonicida experiments were performed as described (Brooks et al 2003;Lillie et al 2006;Young et al 2007), with minor changes. Briefly, 5 mL cultures of A. salmonicida were grown to an OD 600nm of approximately 0.8.…”