Humoral immunoglobulins of the white sturgeon, Acipenser transmontanus: Partial characterization of and recognition with monoclonal antibodies

Mark A. Adkison, Bernardo Basurco, Ronald Hedrick

Research output: Contribution to journalArticle

20 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)285-298
Number of pages14
JournalDevelopmental and Comparative Immunology
Volume20
Issue number4
DOIs
StatePublished - Jul 1 1996

Fingerprint

Immunoglobulins
Monoclonal Antibodies
Ictaluridae
Bass
Oncorhynchus mykiss
Immunoglobulin Subunits
Molecular Weight
Serum
Serum Globulins
Ion Exchange
Ion Exchange Chromatography
Gel Chromatography
Sequence Analysis
Electrophoresis
Polyacrylamide Gel Electrophoresis
Flow Cytometry
Proteins
Gels
Enzyme-Linked Immunosorbent Assay
Lymphocytes

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 journalArticle

@article{8a8feae63a0848bab1d63271606d7dc5,
title = "Humoral immunoglobulins of the white sturgeon, Acipenser transmontanus: Partial characterization of and recognition with monoclonal antibodies",
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.",
keywords = "Acipenser transmontanus, ELISA, Flow cytometry, IgM, Immunoblotting, Immunoglobulin, Monoclonal antibodies, Sturgeon",
author = "Adkison, {Mark A.} and Bernardo Basurco and Ronald Hedrick",
year = "1996",
month = "7",
day = "1",
doi = "10.1016/0145-305X(96)00015-8",
language = "English (US)",
volume = "20",
pages = "285--298",
journal = "Developmental and Comparative Immunology",
issn = "0145-305X",
publisher = "Elsevier Limited",
number = "4",

}

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 -