ELISA detection of human IgG subclass antibodies to Streptococcus mutans

SUMMARYFive rubella antigens were evaluated in an antiglobulin enzyme-linked immunosorbent assay for rubella-specific IgG subclass antibody. One monoclonal anti-human IgG subclass antibody was used for each of IgG1, IgG2and IgG4, but two were compared for IgG3. A total of 101 sera were tested from cases of rubella in the distant past and from cases of primary rubella, reinfection and following immunization. Only one serum gave a discrepant result for specific IgG1, being positive with only one rubella antigen, a commercially prepared antigen coated on to microtitre wells (Enzygnost; Behringwerke). No sera contained detectable specific IgG2. Only four sera contained specific IgG4, and this was detectable only with Enzygnost antigen. For specific IgG3little difference was observed between the two monoclonal anti-human IgG3subclass antibodies; only two very weakly positive sera gave discrepant results. However, varying results were obtained for specific IgG3with the different antigens. Enzygnost gave more positive results for specific IgG3with most categories of sera.It is concluded that the differences between various reports of the rubella-specific IgG subclass profile cannot be explained entirely by the use of different rubella antigens.

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Immunochemical Characterisation Of Two Human Factor IX Inhibitors And Treatment With Activated Concentrate

10.1055/s-0038-1652369 ◽

1981 ◽

Author(s):

J C Giddings ◽

A L Bloom

Keyword(s):

Exchange Chromatography ◽

Factor Ix ◽

Therapeutic Trial ◽

Igg Subclass ◽

Human Igg ◽

Heavy Chains ◽

Human Factor Ix ◽

Activity Unit ◽

Factor Ix Deficiency

Plasma was obtained from two unrelated patients (A and B) with severe hereditary factor IX deficiency who had developed specific inhibitors of factor IX. Immunoglobulin (Ig) was fractionated by salt precipitation and ion exchange chromatography and resulted in 90% recovery of the factor IX inhibitor. Inhibitor plasma or Ig fractions were mixed with specific antisera to ig chains, incubated and centrifuged. Supernatant was assayed for residual inhibitor. Antisera, specific for the heavy chains of human IgG, IgA, IgM, IgD and IgE and the subclasses 1 to 4 of human IgG were used together with antisera specific for the kappa and lambda light chains of IgG. Results of inhibitor neutralisation assays were the same whether plasma or Ig fractions were used. Inhibitor from patient A was completely neutralised by anti IgG and by anti-lambda serum, and was largely neutralised by antiserum to IgG subclass 4. Other antisera had no effect. Inhibitor from patient B was completely neutralised by anti IgG and partially neutralised by anti-kappa and anti-lambda serum. Partial neutralisation of inhibitor was also seen with antisera to IgG subclass 1 and IgG subclass 4. Other antisera again had no effect. Inhibitor A inactivated factor IX instantaneously to a titre of 1 in 100. Neither conventional factor IX concentrate (Oxford DE) at 1 u/ml nor activated‘factor IX’ concentrate (Feiba, Immuno) at 1 u/ml (‘activity’ unit) corrected the kaolin cephalin clotting time (KCCT) of patient A in vitro. However after intravenous administration of Feiba (25 u/kg) progressive shortening of the KCCT occurred from 127 sec. to 60 sec. at 60 min. and was followed by rapid resolution of a haemarthrosis. It is concluded that factor IX antibodies may be of monoclonal or oligoclonal immunoglobulin class and that on development, therapeutic trial of activated factor IX-prothrombin concentrates is indicated for the control of bleeding.

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Human IgG subclass assays using a novel assay method

Journal of Immunological Methods ◽

10.1016/0022-1759(86)90052-9 ◽

1986 ◽

Vol 88 (1) ◽

pp. 65-73 ◽

Cited By ~ 5

Author(s):

Joseph Mayus ◽

Kim Macke ◽

Penelope Shackelford ◽

Jerry Kim ◽

Moon Nahm

Keyword(s):

Assay Method ◽

Igg Subclass ◽

Human Igg

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Unexplained Abuses of Human IgG Subclass Denomination in Antibody Patents

BioDrugs ◽

10.1007/s40259-014-0095-0 ◽

2014 ◽

Vol 28 (4) ◽

pp. 327-329

Author(s):

Jérémy Pottier ◽

Hervé Watier

Keyword(s):

Igg Subclass ◽

Human Igg

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A comparative study of the N-linked oligosaccharide structures of human IgG subclass proteins

Biochemical Journal ◽

10.1042/bj2680529 ◽

1990 ◽

Vol 268 (3) ◽

pp. 529-537 ◽

Cited By ~ 113

Author(s):

R Jefferis ◽

J Lund ◽

H Mizutani ◽

H Nakagawa ◽

Y Kawazoe ◽

...

Keyword(s):

Comparative Study ◽

Internal Space ◽

Igg Subclass ◽

Human Igg ◽

Polyclonal Igg ◽

Oligosaccharide Structures

Quantitative oligosaccharide profiles were determined for each of 18 human IgG paraproteins representing the four subclasses. Each paraprotein exhibits a unique profile that may be substantially different from that observed for polyclonal IgG. The IgG2 and some IgG3 proteins analysed exhibit a predominance of oligosaccharide moieties having galactose on the Man(alpha 1----3) arm rather than the Man(alpha 1----6) arm; it was previously held that galactosylation of the Man(alpha 1----6) arm is preferred, as observed for IgG1, IgG4 and polyclonal IgG. An IgG4 protein is reported that has galactosylated Man(alpha 1----3) and Man(alpha 1----6) arms on both Fc-localized carbohydrate moieties; previous findings suggested that such fully glycosylated structures could not be accommodated within the internal space of the C gamma 2 domains. Unusual monoantennary oligosaccharides present in IgG2 and IgG3 proteins were isolated and their structures determined.

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Quantitative and Qualitative Analysis of Human IgG Subclass Specific mRNA Using Solution Hybridization

Scandinavian Journal of Immunology ◽

10.1111/j.1365-3083.1991.tb01579.x ◽

1991 ◽

Vol 34 (5) ◽

pp. 557-564 ◽

Cited By ~ 5

Author(s):

P. SIDERAS ◽

L. NILSSON ◽

K. B. ISLAM ◽

H. ERICSSON ◽

L. HAMMARSTRoM ◽

...

Keyword(s):

Qualitative Analysis ◽

Igg Subclass ◽

Human Igg ◽

Specific Mrna

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Human IgG Allotypes Co-Occurring in More than One IgG Subclass

Vox Sanguinis ◽

10.1159/000465404 ◽

1982 ◽

Vol 43 (6) ◽

pp. 301-309 ◽

Cited By ~ 9

Author(s):

Erna van Loghem ◽

Gerda de Lange ◽

A.M. van Leeuwen ◽

P.H. van Eeda ◽

L.E. Nijenhuis ◽

...

Keyword(s):

Igg Subclass ◽

Human Igg

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FEASIBILITY OF STUDYING THE FUNCTIONAL NATURE OF HUMAN IgG SUBCLASS RESPONSES BY MEANS OF ANALYTICAL ISOTACHOPHORESIS

Electrophoresis '79 ◽

10.1515/9783111713625-070 ◽

1980 ◽

pp. 765-774

Author(s):

K. W. Hedlund ◽

R. Wistar ◽

D. Nichelson

Keyword(s):

Igg Subclass ◽

Human Igg ◽

Functional Nature

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Evaluation of Human IgG Subclass Assays on Beckman Array

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1177/000456329503200304 ◽

1995 ◽

Vol 32 (3) ◽

pp. 281-288 ◽

Cited By ~ 10

Author(s):

M Pressac ◽

F Allouche ◽

R Circaud ◽

P Aymard

Keyword(s):

Lower Limit ◽

Igg Subclasses ◽

Radial Immunodiffusion ◽

Routine Analysis ◽

Igg Subclass ◽

Human Igg ◽

Protein System

Automated immunonephelometric assays were developed to measure human IgG subclasses in serum, on the Beckman Array Protein System (APSR), with sheep antihuman IgG subclass antisera. The interassay imprecision was judged to be satisfactory in each case (CV: IgG,: 2·3%; IgG2: 2·9%; IgG3: 4·0% and IgG4 5·4%). The standard curves were found to be linear in the ranges 1·5–30·0 g/L for IgG1, 1·5–15·0 g/L for IgG2, 0·1–1·0 g/L for IgG3 and 0·1–1·0 g/L for IgG4. The lower limit for quantification of the immunonephelometric assays was 0·50 g/L for IgG1, 0·04 g/L for IgG2, 0·06 g/L for IgG3 and 0·03 g/L for IgG4. We found no antigen excess to at least 60·0 g/L for IgG, and IgG2, 3·5 g/L for IgG3 and 2·0 g/L for IgG4. Nephelometric results correlated with those of radial immunodiffusion (r = 0·96 for IgG1, r = 0·93 for IgG2, r = 0·90 for IgG3 and r = 0·96 for IgG4). The reaction monitored by the Beckman Array Protein System is easy to perform, rapid and precise. The performance makes the immunonephelometric method suitable for the determination of IgG subclass concentrations in routine analysis.

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