Anti-immunoglobulin antisera used in an ELISA to detect antibodies in barramundi Lates calcarifer to Cryptocaryon irritans.
نویسندگان
چکیده
Immunoglobulins (Ig) in serum from barramundi vaccinated with bovine serum albumin (BSA) were purified by ammonium sulphate precipitation and affinity chromatography using BSA as the ligand. The BSA-binding activity of eluted putative Ig fractions was assessed by enzyme-linked immunosorbent assay (ELISA) before being pooled and characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Double affinity purification did not improve the purity of the Ig preparation compared to single affinity purification. Barramundi Ig were injected into sheep to produce anti-Ig antisera which were assessed in an indirect ELISA as the secondary antibody to detect serum Ig in barramundi vaccinated with Cryptocaryon irritans theronts. Affinity-purified Ig induced a more specific reagent for use as secondary antibody in ELISA than did normal whole-barramundi sera. The heavy (H) chain of barramundi Ig had an apparent molecular weight of 70 kDa while that of the light (L) chain was 27 kDa in SDS-PAGE studies. Under non-reducing conditions 2 putative populations of Ig were identified, at 768 and 210 kDa. The N-terminal sequence of the barramundi Ig H chain showed 78% homology with channel catfish Ictalurus punctatus Ig H chain sequence.
منابع مشابه
The use of biomarkers in barramundi (Lates calcarifer) to monitor contaminants in estuaries of Tropical North Queensland
متن کامل
Effects of modified atmosphere packaging on microbiological load and physico-chemical properties of barramundi (Lates calcarifer Bloch) fillets at 8°C
Different modified atmosphere conditions with various CO2 concentrations on microbial load (psychrophile, mesophile aerobic and anaerobic bacteria) and physico-chemical properties of barramundi (Lates calcarifer Bloch) fillets stored at 8°C were compared to determine the best packaging conditions. The gas conditions evaluated were 100% CO2 (M1), 75% CO2/25% N2 (M2), 50% CO2/50% N2 (M3), 25% CO...
متن کاملAcute and postexposure effects of ammonia toxicity on juvenile barramundi (Lates calcarifer [Bloch]).
Lethal and sublethal effects of ammonia toxicity to juvenile barramundi (Lates calcarifer) were investigated under laboratory conditions following the OECD guidelines for testing of chemicals. Acute toxicity was tested in a static nonrenewal system at pH 9.0 and temperature around 29 degrees C. The 24-, 48-, and 96-h LC(50) values for barramundi were 3.89, 3.67, and 3.31 mg total ammonia N L(-1...
متن کاملEffects of modified atmosphere packaging on microbiological load and physico-chemical properties of barramundi (Lates calcarifer Bloch) fillets at 8°C
Different modified atmosphere conditions with various CO2 concentrations on microbial load (psychrophile, mesophile aerobic and anaerobic bacteria) and physico-chemical properties of barramundi (Lates calcarifer Bloch) fillets stored at 8°C were compared to determine the best packaging conditions. The gas conditions evaluated were 100% CO2 (M1), 75% CO2/25% N2 (M2), 50% CO2/50% N2 (M3), 25% CO...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Diseases of aquatic organisms
دوره 36 1 شماره
صفحات -
تاریخ انتشار 1999