Quantification of lectin in freshwater prawn (Macrobrachium rosenbergii ) hemolymph by ELISA.
An enzyme-linked immunoabsorbent assay was developed to quantify the lectin present in the hemolymph of the freshwater prawn Macrobrachium rosenbergii. This method involves the use of murine monoclonal IgG1 with kappa light chain (designated as 3G1) antibodies raised against the purified lectin, the assay that we developed recognized as little as 30 ng/ml of lectin, and was used to measure the lectin concentration in animals at different maturation stages. The highest concentration of lectin was identified in the hemolymph from post-larval prawns and the lowest in molt stage adult animals. The hemagglutination activity of the lectin was four-fold higher in adult than in juvenile specimens, although in all cases N-acetylated sugar residues, such as N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, and N-acetyl-D-neuraminic acid were inhibitors of the lectin activity, suggesting that lectin plays a role in the transport of N-acetylated sugar in juvenile prawns. Our results indicate that lectin concentration and hemagglutinating activity could be influenced by developmental conditions of the freshwater prawn.
Agundis
C
C
Departamento de Bioquimica, Facultad de Medicina UNAM, Mexico City, Mexico. agundis@bq.unam.mx
Pereyra
A
A
Zenteno
R
R
Brassart
C
C
Sierra
C
C
Vazquez
L
L
Zenteno
E
E
D000116:Acetylgalactosamine / Q000494:pharmacology; D000117:Acetylglucosamine / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006458:Hemolymph / Q000378:metabolism; D006825:Hybridomas / Q000378:metabolism; D037102:Lectins / Q000032:analysis* / Q000378:metabolism* / Q000502:physiology; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000494:pharmacology; D033541:Palaemonidae; D013997:Time Factors
D000911:Antibodies, Monoclonal; D037102:Lectins; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine; D000117:Acetylglucosamine