Purification and characterization of IgM-like immunoglobulin from turbot (Scophthalmus maximus L.).
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Purification and characterization of IgM-like immunoglobulin from turbot (Scophthalmus maximus L.). / Kofod, H.; Pedersen, K.; Larsen, J. L.; Buchmann, K.
I: Acta Veterinaria Scandinavica, Bind 35, Nr. 1, 1994, s. 1-10.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Purification and characterization of IgM-like immunoglobulin from turbot (Scophthalmus maximus L.).
AU - Kofod, H.
AU - Pedersen, K.
AU - Larsen, J. L.
AU - Buchmann, K.
PY - 1994
Y1 - 1994
N2 - A total of 40 turbot (Scophthalmus maximus) were immunized 3 times during a 3 months period using DNP-HSA whereafter serum samples were collected and pooled. Specific immunoglobulins (Ig) were affinity purified on an agarose column with immobilized DNP-BSA and further purified by gel filtration whereafter monospecific rabbit anti Ig serum was generated. Size exclusion chromatography and non-reduced SDS-PAGE indicated a MW of 8-900 kDa of the dominant antigen binding proteins from turbot serum. Reduced SDS-PAGE showed this fraction to be composed of disulphide linked heavy and light chains with MWs of 79 and 27-29 kDa, respectively, indicating a tetrameric structure. Isoelectric focusing of the 800-900 kDa Ig showed several bands between pH 5.5 and pH 5.8. Mean Ig concentration in serum of 10 turbot was measured to 6.48 mg/ml (SD 5.4) using rocket immunoelectrophoresis. Low molecular weight antigen binding molecules were copurified with the dominating immunoglobulins with an estimated MW of 500 kDa. Reducing SDS-PAGE of this fraction revealed molecules with MWs of 97, 79, 57, 29, and 27 kDa.
AB - A total of 40 turbot (Scophthalmus maximus) were immunized 3 times during a 3 months period using DNP-HSA whereafter serum samples were collected and pooled. Specific immunoglobulins (Ig) were affinity purified on an agarose column with immobilized DNP-BSA and further purified by gel filtration whereafter monospecific rabbit anti Ig serum was generated. Size exclusion chromatography and non-reduced SDS-PAGE indicated a MW of 8-900 kDa of the dominant antigen binding proteins from turbot serum. Reduced SDS-PAGE showed this fraction to be composed of disulphide linked heavy and light chains with MWs of 79 and 27-29 kDa, respectively, indicating a tetrameric structure. Isoelectric focusing of the 800-900 kDa Ig showed several bands between pH 5.5 and pH 5.8. Mean Ig concentration in serum of 10 turbot was measured to 6.48 mg/ml (SD 5.4) using rocket immunoelectrophoresis. Low molecular weight antigen binding molecules were copurified with the dominating immunoglobulins with an estimated MW of 500 kDa. Reducing SDS-PAGE of this fraction revealed molecules with MWs of 97, 79, 57, 29, and 27 kDa.
UR - http://www.scopus.com/inward/record.url?scp=0028252315&partnerID=8YFLogxK
M3 - Journal article
C2 - 8209813
AN - SCOPUS:0028252315
VL - 35
SP - 1
EP - 10
JO - Acta Veterinaria Scandinavica, Supplement
JF - Acta Veterinaria Scandinavica, Supplement
SN - 0065-1699
IS - 1
ER -
ID: 247152078