Isoelectric focusing-affinity immunoblot analysis of mouse monoclonal antibodies to the four human IgG subclasses

1987 ◽  
Vol 8 (3) ◽  
pp. 127-134 ◽  
Author(s):  
Robert G. Hamilton ◽  
Marianne Roebber ◽  
Charles B. Reimer ◽  
L. Scott Rodkey
Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1475-1480 ◽  
Author(s):  
CA Fulcher ◽  
S de Graaf Mahoney ◽  
TS Zimmerman

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.


Blood ◽  
1987 ◽  
Vol 69 (5) ◽  
pp. 1475-1480 ◽  
Author(s):  
CA Fulcher ◽  
S de Graaf Mahoney ◽  
TS Zimmerman

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.


1987 ◽  
Vol 33 (10) ◽  
pp. 1707-1725 ◽  
Author(s):  
R G Hamilton

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.


1992 ◽  
Vol 31 (2) ◽  
pp. 143-168 ◽  
Author(s):  
R. Jefferis ◽  
C.B. Reimer ◽  
F. Skvaril ◽  
G.G. de Lange ◽  
D.M. Goodall ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document