Abstract
White sturgeon (Acipenser transmontanus) immunoglobulin (Ig) was purified from serum by two methods, ion-exchange chromatography and gel filtration and precipitation of the euglobulin fraction. The purity of these immunoglobulin preparations was confirmed by gel electrophoresis. Sequence analysis of the N-terminal amino acids confirmed that the purified protein was immunoglobulin. The major portion of the immunoglobulin preparation consisted of two proteins with estimated molecular weights (m.w.) of 870 and 170 kDa. The m.w. of the H- and L-chains of the purified Ig were 73 and 27- 30 kDa, respectively, as determined by SDS-PAGE. Ion-exchange purified Ig was used to immunize mice for the production of monoclonal antibodies. This resulted in the production of six stable hybrids that recognized sturgeon Ig, two specific for heavy chain and four specific for light chain. The two anti- H-chain mabs were highly specific for white sturgeon Ig while all four anti- L-chain mabs cross reacted with Ig from green sturgeon (A. medirostris), Atlantic sturgeon (A. oxyrhynchus oxyrhynchus), shovelnose sturgeon (Scaphirhynchus platorynchus), and paddlefish (Polyodon spathula), (all Chondrosteans), but not with channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss) or striped bass (Morone saxatilis). Themabs were used to enumerate the percentage of sIg+ lymphocytes in the peripheral blood of white sturgeon by flow cytometry. The percentage of cells positively stained with the mabs ranged from 12 to 28%. In a comparison of mabs with polyclonal rabbit anti-sturgeon Ig serum by ELISA the mabs produced a larger signal and less background than the polyclonal serum.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 285-298 |
| Number of pages | 14 |
| Journal | Developmental and Comparative Immunology |
| Volume | 20 |
| Issue number | 4 |
| DOIs | |
| State | Published - Jul 1 1996 |
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Keywords
- Acipenser transmontanus
- ELISA
- Flow cytometry
- IgM
- Immunoblotting
- Immunoglobulin
- Monoclonal antibodies
- Sturgeon
ASJC Scopus subject areas
- Immunology
- Developmental Biology
Cite this
Humoral immunoglobulins of the white sturgeon, Acipenser transmontanus : Partial characterization of and recognition with monoclonal antibodies. / Adkison, Mark A.; Basurco, Bernardo; Hedrick, Ronald.
In: Developmental and Comparative Immunology, Vol. 20, No. 4, 01.07.1996, p. 285-298.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Humoral immunoglobulins of the white sturgeon, Acipenser transmontanus
T2 - Partial characterization of and recognition with monoclonal antibodies
AU - Adkison, Mark A.
AU - Basurco, Bernardo
AU - Hedrick, Ronald
PY - 1996/7/1
Y1 - 1996/7/1
N2 - White sturgeon (Acipenser transmontanus) immunoglobulin (Ig) was purified from serum by two methods, ion-exchange chromatography and gel filtration and precipitation of the euglobulin fraction. The purity of these immunoglobulin preparations was confirmed by gel electrophoresis. Sequence analysis of the N-terminal amino acids confirmed that the purified protein was immunoglobulin. The major portion of the immunoglobulin preparation consisted of two proteins with estimated molecular weights (m.w.) of 870 and 170 kDa. The m.w. of the H- and L-chains of the purified Ig were 73 and 27- 30 kDa, respectively, as determined by SDS-PAGE. Ion-exchange purified Ig was used to immunize mice for the production of monoclonal antibodies. This resulted in the production of six stable hybrids that recognized sturgeon Ig, two specific for heavy chain and four specific for light chain. The two anti- H-chain mabs were highly specific for white sturgeon Ig while all four anti- L-chain mabs cross reacted with Ig from green sturgeon (A. medirostris), Atlantic sturgeon (A. oxyrhynchus oxyrhynchus), shovelnose sturgeon (Scaphirhynchus platorynchus), and paddlefish (Polyodon spathula), (all Chondrosteans), but not with channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss) or striped bass (Morone saxatilis). Themabs were used to enumerate the percentage of sIg+ lymphocytes in the peripheral blood of white sturgeon by flow cytometry. The percentage of cells positively stained with the mabs ranged from 12 to 28%. In a comparison of mabs with polyclonal rabbit anti-sturgeon Ig serum by ELISA the mabs produced a larger signal and less background than the polyclonal serum.
AB - White sturgeon (Acipenser transmontanus) immunoglobulin (Ig) was purified from serum by two methods, ion-exchange chromatography and gel filtration and precipitation of the euglobulin fraction. The purity of these immunoglobulin preparations was confirmed by gel electrophoresis. Sequence analysis of the N-terminal amino acids confirmed that the purified protein was immunoglobulin. The major portion of the immunoglobulin preparation consisted of two proteins with estimated molecular weights (m.w.) of 870 and 170 kDa. The m.w. of the H- and L-chains of the purified Ig were 73 and 27- 30 kDa, respectively, as determined by SDS-PAGE. Ion-exchange purified Ig was used to immunize mice for the production of monoclonal antibodies. This resulted in the production of six stable hybrids that recognized sturgeon Ig, two specific for heavy chain and four specific for light chain. The two anti- H-chain mabs were highly specific for white sturgeon Ig while all four anti- L-chain mabs cross reacted with Ig from green sturgeon (A. medirostris), Atlantic sturgeon (A. oxyrhynchus oxyrhynchus), shovelnose sturgeon (Scaphirhynchus platorynchus), and paddlefish (Polyodon spathula), (all Chondrosteans), but not with channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss) or striped bass (Morone saxatilis). Themabs were used to enumerate the percentage of sIg+ lymphocytes in the peripheral blood of white sturgeon by flow cytometry. The percentage of cells positively stained with the mabs ranged from 12 to 28%. In a comparison of mabs with polyclonal rabbit anti-sturgeon Ig serum by ELISA the mabs produced a larger signal and less background than the polyclonal serum.
KW - Acipenser transmontanus
KW - ELISA
KW - Flow cytometry
KW - IgM
KW - Immunoblotting
KW - Immunoglobulin
KW - Monoclonal antibodies
KW - Sturgeon
UR - http://www.scopus.com/inward/record.url?scp=0030199460&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0030199460&partnerID=8YFLogxK
U2 - 10.1016/0145-305X(96)00015-8
DO - 10.1016/0145-305X(96)00015-8
M3 - Article
C2 - 8915630
AN - SCOPUS:0030199460
VL - 20
SP - 285
EP - 298
JO - Developmental and Comparative Immunology
JF - Developmental and Comparative Immunology
SN - 0145-305X
IS - 4
ER -