Preparation of Anti-GM4 antiserum and its assay by a solid-phase radioimmunoassay

Ronald I. Jacobson; Noriyuki Kasai; Frank F. Richards; Robert K. Yu

doi:10.1016/0165-5728(82)90025-X

Preparation of Anti-G_M4 antiserum and its assay by a solid-phase radioimmunoassay

Ronald I. Jacobson, Noriyuki Kasai, Frank F. Richards, Robert K. Yu

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

Anti-G_M4 antiserum was prepared from rabbits by immunization with pure human brain G_M4 ganglioside in complete Freund's adjuvant and methylated bovine serum albumin. None of the immunized animals developed any clinically apparent neurological dysfunction. The antiserum titer and specificity were analyzed by double immunodiffusion and a novel solid-phase radioimmunoassay (RIA). In the latter procedure, microtiter plate wells were coated first with the glycolipid antigen, followed by sequential application of antiserum and [¹²⁵I]-Staphylococcal Protein A. The absorbed radioactivity in the well was then counted. Employing the RIA procedure, anti-G_M4 antibody achieved a titer of 1:1600. The antiserum also exhibited a high degree of specificity to G_M4; cross-reactivity with glycolipids of similar structure was negligible. The production of highly specific antiserum to G_M4 and the feasibility of detecting antibodies to glycolipid antigens by a convenient solid-phase RIA should be useful to further study the biological and immunological roles of G_M4 and other glycolipids in the central nervous system.

Original language	English (US)
Pages (from-to)	225-235
Number of pages	11
Journal	Journal of Neuroimmunology
Volume	3
Issue number	3
DOIs	https://doi.org/10.1016/0165-5728(82)90025-X
State	Published - Nov 1982
Externally published	Yes

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Neurology
Clinical Neurology

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10.1016/0165-5728(82)90025-X

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@article{2c4c3bc125e649148179174dfb4f86d3,

title = "Preparation of Anti-GM4 antiserum and its assay by a solid-phase radioimmunoassay",

abstract = "Anti-GM4 antiserum was prepared from rabbits by immunization with pure human brain GM4 ganglioside in complete Freund's adjuvant and methylated bovine serum albumin. None of the immunized animals developed any clinically apparent neurological dysfunction. The antiserum titer and specificity were analyzed by double immunodiffusion and a novel solid-phase radioimmunoassay (RIA). In the latter procedure, microtiter plate wells were coated first with the glycolipid antigen, followed by sequential application of antiserum and [125I]-Staphylococcal Protein A. The absorbed radioactivity in the well was then counted. Employing the RIA procedure, anti-GM4 antibody achieved a titer of 1:1600. The antiserum also exhibited a high degree of specificity to GM4; cross-reactivity with glycolipids of similar structure was negligible. The production of highly specific antiserum to GM4 and the feasibility of detecting antibodies to glycolipid antigens by a convenient solid-phase RIA should be useful to further study the biological and immunological roles of GM4 and other glycolipids in the central nervous system.",

author = "Jacobson, {Ronald I.} and Noriyuki Kasai and Richards, {Frank F.} and Yu, {Robert K.}",

note = "Funding Information: This work wa~i supported by USPH$ grants NS+~I853, AI 08614 and NS 0f~032, and a gtaat from the National Multiple Sclerosis Society (RG 1289 A 2). Address reprhtt requests and correspondencet o ~Y. The present address o!~R IJ is DiC'tsioao f l~a~ Neurolosy, ~tox 486 Mayo, University of Minnesota School of l~¢dicine, Minneapolis. M'b, ~$diS$, U.S.A. * The geaglioside nomenclature system of Sv~lerholm (1%3) is e~ied. Other al~ecatie¢~ used are: NONA, N-glyoolylneuraminica cid; Gal-Cer., 8alactocet'ebroside;S OeGaI.C.~, sullaude.",

year = "1982",

month = nov,

doi = "10.1016/0165-5728(82)90025-X",

language = "English (US)",

volume = "3",

pages = "225--235",

journal = "Journal of Neuroimmunology",

issn = "0165-5728",

publisher = "Elsevier",

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TY - JOUR

T1 - Preparation of Anti-GM4 antiserum and its assay by a solid-phase radioimmunoassay

AU - Jacobson, Ronald I.

AU - Kasai, Noriyuki

AU - Richards, Frank F.

AU - Yu, Robert K.

N1 - Funding Information: This work wa~i supported by USPH$ grants NS+~I853, AI 08614 and NS 0f~032, and a gtaat from the National Multiple Sclerosis Society (RG 1289 A 2). Address reprhtt requests and correspondencet o ~Y. The present address o!~R IJ is DiC'tsioao f l~a~ Neurolosy, ~tox 486 Mayo, University of Minnesota School of l~¢dicine, Minneapolis. M'b, ~$diS$, U.S.A. * The geaglioside nomenclature system of Sv~lerholm (1%3) is e~ied. Other al~ecatie¢~ used are: NONA, N-glyoolylneuraminica cid; Gal-Cer., 8alactocet'ebroside;S OeGaI.C.~, sullaude.

PY - 1982/11

Y1 - 1982/11

N2 - Anti-GM4 antiserum was prepared from rabbits by immunization with pure human brain GM4 ganglioside in complete Freund's adjuvant and methylated bovine serum albumin. None of the immunized animals developed any clinically apparent neurological dysfunction. The antiserum titer and specificity were analyzed by double immunodiffusion and a novel solid-phase radioimmunoassay (RIA). In the latter procedure, microtiter plate wells were coated first with the glycolipid antigen, followed by sequential application of antiserum and [125I]-Staphylococcal Protein A. The absorbed radioactivity in the well was then counted. Employing the RIA procedure, anti-GM4 antibody achieved a titer of 1:1600. The antiserum also exhibited a high degree of specificity to GM4; cross-reactivity with glycolipids of similar structure was negligible. The production of highly specific antiserum to GM4 and the feasibility of detecting antibodies to glycolipid antigens by a convenient solid-phase RIA should be useful to further study the biological and immunological roles of GM4 and other glycolipids in the central nervous system.

AB - Anti-GM4 antiserum was prepared from rabbits by immunization with pure human brain GM4 ganglioside in complete Freund's adjuvant and methylated bovine serum albumin. None of the immunized animals developed any clinically apparent neurological dysfunction. The antiserum titer and specificity were analyzed by double immunodiffusion and a novel solid-phase radioimmunoassay (RIA). In the latter procedure, microtiter plate wells were coated first with the glycolipid antigen, followed by sequential application of antiserum and [125I]-Staphylococcal Protein A. The absorbed radioactivity in the well was then counted. Employing the RIA procedure, anti-GM4 antibody achieved a titer of 1:1600. The antiserum also exhibited a high degree of specificity to GM4; cross-reactivity with glycolipids of similar structure was negligible. The production of highly specific antiserum to GM4 and the feasibility of detecting antibodies to glycolipid antigens by a convenient solid-phase RIA should be useful to further study the biological and immunological roles of GM4 and other glycolipids in the central nervous system.

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Preparation of Anti-G_M4 antiserum and its assay by a solid-phase radioimmunoassay

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