Preparation and characterization of antibody to galactosyl( α1 → 4)galactosyl( β1 → 4)glucosylceramide
Antiserum against galactosyl( α1 → 4)galactosyl( β1 → 4)glucosylceramide (globotriaosylceramide, Gb 3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycolipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb...
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| Published in | Biochimica et biophysica acta Vol. 582; no. 2; pp. 283 - 294 |
|---|---|
| Main Authors | , |
| Format | Journal Article |
| Language | English |
| Published |
Netherlands
Elsevier B.V
18.01.1979
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0304-4165 0006-3002 1872-8006 |
| DOI | 10.1016/0304-4165(79)90391-X |
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| Abstract | Antiserum against galactosyl(
α1 → 4)galactosyl(
β1 → 4)glucosylceramide (globotriaosylceramide, Gb
3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycolipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb
3 activity was quantitated initially by immunoprecipitation employing Gb
3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb
3/lecithin/cholesterol (1 : 6 : 20, by wt.) showed antiGb
3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with sphingomyelin/cholesterol/dicetylphosphate/Gb
3 (molar ratio, 100 : 75 : 11 : 1). The antibody crossreacted with the trisaccharide, Gal(
α1 → 4)Gal(
β1 → 4)Glc, but not with galactosylceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides. |
|---|---|
| AbstractList | Antiserum against galactosyl(
α1 → 4)galactosyl(
β1 → 4)glucosylceramide (globotriaosylceramide, Gb
3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycolipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb
3 activity was quantitated initially by immunoprecipitation employing Gb
3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb
3/lecithin/cholesterol (1 : 6 : 20, by wt.) showed antiGb
3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with sphingomyelin/cholesterol/dicetylphosphate/Gb
3 (molar ratio, 100 : 75 : 11 : 1). The antibody crossreacted with the trisaccharide, Gal(
α1 → 4)Gal(
β1 → 4)Glc, but not with galactosylceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides. Antiserum against galactosyl(alpha 1 leads to 4)galactosyl(beta 1 leads to 4)glucosylceramide (globotriaosylceramide, Gb3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycoplipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb3 activity was quantitated initially by immunoprescipitation employing Gb3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb3/lecithin/cholesterol (1 :6 :20, by wt.) showed antiGb3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with shingomyelin/cholesterol/dicetylphosphate/Gb3 (molar ratio, 100 : 75 :11 :1). The antibody crossreacted with the trisaccharide, Gal(alpha1 leads to 4)Gal(beta1 leads to 4)Glc, but not with galactosylerceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides. Antiserum against galactosyl(alpha 1 leads to 4)galactosyl(beta 1 leads to 4)glucosylceramide (globotriaosylceramide, Gb3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycoplipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb3 activity was quantitated initially by immunoprescipitation employing Gb3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb3/lecithin/cholesterol (1 :6 :20, by wt.) showed antiGb3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with shingomyelin/cholesterol/dicetylphosphate/Gb3 (molar ratio, 100 : 75 :11 :1). The antibody crossreacted with the trisaccharide, Gal(alpha1 leads to 4)Gal(beta1 leads to 4)Glc, but not with galactosylerceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides.Antiserum against galactosyl(alpha 1 leads to 4)galactosyl(beta 1 leads to 4)glucosylceramide (globotriaosylceramide, Gb3) was raised in rabbits by the administration of four weekly intramuscular injections of 1.5 mg of the purified glycoplipid along with bovine serum albumin and Freund's complete adjuvant. AntiGb3 activity was quantitated initially by immunoprescipitation employing Gb3 mixed with 100-fold excess of lecithin and cholesterol (1 : 1 or 1 : 2, by wt.) as antigen. Subsequently, complement fixation tests done with antigen preparations containing Gb3/lecithin/cholesterol (1 :6 :20, by wt.) showed antiGb3 titres of up to 1 : 8192. Fractionation of the antiserum by BioGel A5m chromatography indicated the antibody was an IgM immunoglobulin. The partially purified antibody stimulated complement-dependent release of glucose from glucose-containing liposomes prepared with shingomyelin/cholesterol/dicetylphosphate/Gb3 (molar ratio, 100 : 75 :11 :1). The antibody crossreacted with the trisaccharide, Gal(alpha1 leads to 4)Gal(beta1 leads to 4)Glc, but not with galactosylerceramide, lactosylceramide, GM1 ganglioside, globotetraosylceramide, digalactosyldiglyceride or a number of low molecular weight saccharides. |
| Author | Embil, J.A. Clarke, J.T.R. |
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| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/760826$$D View this record in MEDLINE/PubMed |
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| Cites_doi | 10.1016/0022-1759(75)90078-2 10.1016/0019-2791(74)90118-9 10.1016/0006-291X(71)90704-2 10.1016/S0065-2776(08)60321-1 10.1016/0019-2791(68)90181-X 10.1016/0005-2760(76)90152-1 10.1021/bi00693a010 10.1016/S0022-2275(20)36996-0 10.1016/0005-2760(76)90289-7 10.4049/jimmunol.104.6.1530 10.1016/S0021-9258(18)61846-0 10.1126/science.177.4053.949 10.1111/j.1423-0410.1969.tb04775.x 10.1016/S0021-9258(18)62195-7 10.1016/0006-291X(77)91395-X 10.1093/oxfordjournals.jbchem.a127880 10.1016/S0021-9258(19)77217-2 10.1016/S0022-2275(20)38884-2 10.1021/bi00838a036 10.1016/S0021-9258(18)51888-3 10.1016/0006-291X(74)90426-4 10.1016/0005-2760(66)90012-9 10.1016/S0021-9258(19)43774-5 |
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| Keywords | Gb 3, Gal( α1 → 4)Gal( β1 → 4)Glc → ceramide (globotriaosylceramide) Glycosphingolipid globotetraosylceramide, GalNAc( β1 → 3)Gal( α1 → 4)Glc → ceramide Liposome Complement fixation GM1 ganglioside, Gal β1 → 3GalNAc β1 → 4(NAcNeu α2 → 3)Gal β1 → 4Glc → ceramide GM3 ganglioside, NAcNeu( α2 → 3) Gal( β1 → 4)Glc → ceramide Globotriaose antibody |
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| Snippet | Antiserum against galactosyl(
α1 → 4)galactosyl(
β1 → 4)glucosylceramide (globotriaosylceramide, Gb
3) was raised in rabbits by the administration of four... Antiserum against galactosyl(alpha 1 leads to 4)galactosyl(beta 1 leads to 4)glucosylceramide (globotriaosylceramide, Gb3) was raised in rabbits by the... |
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| SubjectTerms | Animals Antibodies - isolation & purification Antibody Specificity Cell Membrane - metabolism Complement fixation Cross Reactions Globosides - immunology Globosides - metabolism Globotriaose antibody Glycosphingolipid Glycosphingolipids - immunology Immunochemistry Immunoglobulin M Liposome Rabbits |
| Title | Preparation and characterization of antibody to galactosyl( α1 → 4)galactosyl( β1 → 4)glucosylceramide |
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