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J Vet Sci. 2006 Sep;7(3):293-295 |
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Production of monoclonal antibodies against serum immunoglobulins of black rockfish (Sebastes schlegeli Higendorf)
Geewook Shin1, Hyungjun Lee1, K. J. Palaksha1, Youngrim Kim1, Eunyoung Lee1, Yongseung Shin1, Eunggoo Lee1, Kyungdae Park2, Taesung Jung1,* |
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1Laboratory of Fish & Shellfish Diseases, College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Korea. jungts@gsnu.ac.kr
2Gyeongsangnam-do Fisheries Resources Research Institute, Tongyeong 650-947, Korea |
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The present study was undertaken to produce monoclonal
antibodies (MAbs) against immunoglobulin (Ig) purified
from black rockfish (Sebastes schlegeli Higendorf) serum
using protein A, mannan binding protein, and goat IgG
affinity columns. These three different ligands were found
to possess high affinity for black rockfish serum Ig. All of
the Igs purified eluted at only 0.46 M NaCl concentration
in anion exchange column chromatography and consisted
of two bands at 70 kDa and 25 kDa in SDS-PAGE; they
also had similar antigenicity for MAbs to Ig heavy chain
in immunoblot assays. Therefore, black rockfish Ig is
believed to exist as a single isotype within serum. The MAbs
produced against Ig heavy chain reacted specifically with
spots distributed over the pI range from 4.8 to 5.6 with a
molecular weight of 70 kDa on two dimensional gel
electrophoresis immunoblot profiles.
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