scholarly journals Evaluation of Human IgG Subclass Assays on Beckman Array

1995 ◽  
Vol 32 (3) ◽  
pp. 281-288 ◽  
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.

scholarly journals FVIII inhibitor IgG subclass and FVIII polypeptide specificity determined by immunoblotting

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1475-1480 ◽  
Author(s):  
CA Fulcher ◽  
S de Graaf Mahoney ◽  
TS Zimmerman
Keyword(s):  
Monoclonal Antibodies ◽  
Factor Viii ◽  
Igg Subclasses ◽  
Radial Immunodiffusion ◽  
Igg Subclass ◽  
Human Igg ◽  
Immunoglobulin Class ◽  
Fviii Inhibitor

Abstract We used immunoblotting of purified factor VIII coagulant protein (FVIII) to localize FVIII inhibitor epitopes in 76 inhibitor plasmas to either the 92-kd FVIII polypeptide (and its 54-kd and/or 44-kd thrombin fragments), the 80-kd polypeptide (and its 72-kd thrombin fragment), or both of these polypeptides. We also used immunoblotting to examine the immunoglobulin class and subclass content of 12 inhibitors with monoclonal antibodies specific for human IgG subclasses and IgM. Seven hemophilic (alloantibody) and five spontaneous (autoantibody) inhibitors contained IgG-1 and IgG-4 antibody; one of the spontaneous inhibitors also contained IgG-3. In one hemophilic inhibitor, the IgG-4 component reacted strongly with the 92-kd and 80-kd polypeptides, whereas the IgG-1 component reacted only minimally with the 92-kd polypeptide although its reactivity with the 80-kd polypeptide was strong. Another hemophilic inhibitor was affinity purified and subjected to quantitative radial immunodiffusion, and the presence of IgG-1 and IgG-4 antibody was confirmed. We conclude that the inhibitors examined are not monoclonal but are probably of restricted polyclonal origin and that different IgG subclasses in an inhibitor plasma can have different degrees of FVIII polypeptide reactivity.

scholarly journals FVIII inhibitor IgG subclass and FVIII polypeptide specificity determined by immunoblotting

Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1475-1480 ◽  
Author(s):  
CA Fulcher ◽  
S de Graaf Mahoney ◽  
TS Zimmerman
Keyword(s):  
Monoclonal Antibodies ◽  
Factor Viii ◽  
Igg Subclasses ◽  
Radial Immunodiffusion ◽  
Igg Subclass ◽  
Human Igg ◽  
Immunoglobulin Class ◽  
Fviii Inhibitor

We used immunoblotting of purified factor VIII coagulant protein (FVIII) to localize FVIII inhibitor epitopes in 76 inhibitor plasmas to either the 92-kd FVIII polypeptide (and its 54-kd and/or 44-kd thrombin fragments), the 80-kd polypeptide (and its 72-kd thrombin fragment), or both of these polypeptides. We also used immunoblotting to examine the immunoglobulin class and subclass content of 12 inhibitors with monoclonal antibodies specific for human IgG subclasses and IgM. Seven hemophilic (alloantibody) and five spontaneous (autoantibody) inhibitors contained IgG-1 and IgG-4 antibody; one of the spontaneous inhibitors also contained IgG-3. In one hemophilic inhibitor, the IgG-4 component reacted strongly with the 92-kd and 80-kd polypeptides, whereas the IgG-1 component reacted only minimally with the 92-kd polypeptide although its reactivity with the 80-kd polypeptide was strong. Another hemophilic inhibitor was affinity purified and subjected to quantitative radial immunodiffusion, and the presence of IgG-1 and IgG-4 antibody was confirmed. We conclude that the inhibitors examined are not monoclonal but are probably of restricted polyclonal origin and that different IgG subclasses in an inhibitor plasma can have different degrees of FVIII polypeptide reactivity.

Nephelometry of human IgG subclass concentration in serum.

1981 ◽  
Vol 27 (2) ◽  
pp. 310-313 ◽  
Author(s):  
O E Beck ◽  
P E Kaiser
Keyword(s):  
Human Serum ◽  
Radial Immunodiffusion ◽  
Igg Subclass ◽  
Human Igg ◽  
Specific Antisera ◽  
Igg1 Subclass ◽  
The Individual

Abstract An immunoassay method for determination of human IgG subclass concentration by rate nephelometry was developed by using the Beckman Immunochemistry Analyzer, subclass-specific antisera, and a human serum standard. Twelve sera derived from normal as well as gammapathological patients were analyzed. The total IgG concentration, as determined by using a readily available kit, correlated with the sum of the concentration of the individual subclasses. Most of the gammapathological cases were shown to be the IgG1 subclass, which was confirmed by radial immunodiffusion.

Human IgG subclass measurements in the clinical laboratory.

1987 ◽  
Vol 33 (10) ◽  
pp. 1707-1725 ◽  
Author(s):  
R G Hamilton
Keyword(s):  
Monoclonal Antibodies ◽  
Clinical Laboratory ◽  
Placental Transfer ◽  
Clinical Importance ◽  
Igg Subclasses ◽  
Cell Surface Receptor ◽  
Immunological Methods ◽  
Igg Subclass ◽  
Human Igg

Abstract Complement activation, cell surface-receptor binding, blocking activity, and possibly placental transfer are among the biologically important functional differences that have been detected between the four human IgG subclasses by use of polyclonal antisera. In 1985, a IUIS/WHO panel of immunologists, using eight immunological methods, documented the specificity of select monoclonal antibodies for the IgG subclasses. Clinical assays have been developed involving these monoclonal antibodies that allow quantification of the concentration of IgG subclass protein and distribution of the IgG subclass antibodies in human immune responses. This review addresses issues of concern to investigators who are evaluating and (or) developing quantitative human IgG subclass assays in the clinical laboratory. Unique physical (structural) and biological (functional) properties of human IgG subclasses are summarized, with a focus on aspects pertinent to their clinical importance and in vitro quantification. The HP-series monoclonal antibodies with documented specificity are examined within the context of their application to several immunological methods. I describe unique technical aspects of total and antigen-specific IgG-subclass immunoassays involving these monoclonal antibodies. Finally, this report outlines clinical applications and indications for IgG-subclass measurements in the study of human health and disease.

Specific cleavage sites on human IgG subclasses by cruzipain, the major cysteine proteinase from Trypanosoma cruzi

2003 ◽  
Vol 130 (1) ◽  
pp. 23-29 ◽  
Author(s):  
Patricia Berasain ◽  
Carlos Carmona ◽  
Blas Frangione ◽  
Juan José Cazzulo ◽  
Fernando Goñi
Keyword(s):  
Trypanosoma Cruzi ◽  
Cysteine Proteinase ◽  
Igg Subclasses ◽  
Cleavage Sites ◽  
Human Igg

ELISA detection of human IgG subclass antibodies to Streptococcus mutans

1986 ◽  
Vol 87 (1) ◽  
pp. 95-102 ◽  
Author(s):  
S.J. Challacombe ◽  
M. Biggerstaff ◽  
C. Greenall ◽  
D.M. Kemeny
Keyword(s):  
Streptococcus Mutans ◽  
Igg Subclass ◽  
Human Igg
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