Contributions of heavy and light chains of rabbit immunoglobulin G to antibody activity. II. Binding activities of reconstituted immunoglobulins

Biochemistry ◽  
1972 ◽  
Vol 11 (8) ◽  
pp. 1338-1345 ◽  
Author(s):  
Richard G. Painter ◽  
Harvey J. Sage ◽  
Charles Tanford
Keyword(s):  
Immunoglobulin G ◽  
Light Chains ◽  
Antibody Activity ◽  
Rabbit Immunoglobulin

scholarly journals Transient IgA-lambda paraproteinemia during treatment of acute myelomonoblastic leukemia

Blood ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 21-25 ◽  
Author(s):  
B Van Camp ◽  
P Reynaerts ◽  
JP Naets ◽  
J Radl
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Clonal Expansion ◽  
Cellular Level ◽  
Light Chains ◽  
Antibody Activity ◽  
Intensive Chemotherapy ◽  
Bone Marrow Aplasia ◽  
Large Panel ◽  
Common Antigens

Abstract Monoclonal plasma cell proliferation with secretion of IgA-lambda and free lambda light chains during a phase of bone marrow aplasia following intensive chemotherapy was observed in a patient suffering from acute myelomonoblastic leukemia. The clonal expansion and regression was investigated at the cellular level by immunofluorescence using an antiserum against the idiotype of the paraportein. Although a large panel of common antigens was used for testing, no antibody activity of the paraprotein could be demonstrated.

scholarly journals Peroxidase and fluorescein isothiocyanate as antibody markers. A quantitative comparison of two peroxidase conjugates prepared with glutaraldehyde or periodate anda fluorescein conjugate.

1976 ◽  
Vol 24 (9) ◽  
pp. 1017-1025 ◽  
Author(s):  
D M Broorsma ◽  
J G Steefkerk ◽  
N Kors
Keyword(s):  
Enzyme Activity ◽  
Immunoglobulin G ◽  
Fluorescein Isothiocyanate ◽  
Antibody Activity ◽  
Positive Staining ◽  
Passive Hemagglutination ◽  
Agarose Bead ◽  
Cryostat Sections ◽  
Peroxidase Conjugate ◽  
Considerable Loss

Batches of rabbit anti-human immunoglobulin G antibodies were labeled either with horseradish peroxidase, using the two-step glutaraldehyde method or the periodate method, or with fluorescein isothiocyanate (FITC). The peroxidase conjugates were isolated by chromatography using two different gel types. The five types of conjugates thus obtained were standardized to the same amount of rabbit immunoglobulin G. The antibody activity, as estimated by means of single radial immunodiffusion and passive hemagglutination, and the enzyme activity, determined with orthodianisidine, were compared. The ultimate dilutions and absolute amounts of the five conjugates giving positive reactions were determined in direct and indirect immunohistochemical tests, using both cryostat sections of skin and the agarose bead model system. It appeared that during the peroxidase conjugation procedures there was a considerable loss of abtibody and enzyme activity, whereas in the FITC conjugation procedure the antibody activity remained intact. Neverthe less, peroxidase conjugates prepared with glutaraldehyde still gave positive staining reactions in equal or somewhat higher dilutions than the fluorescin conjugate did. The peroxidase conjugates prepared with periodate could not be diluted to the same extent. For the detection of antibodies by indirect immunohistochemical methods, the peroxidase conjugate, prepared with glutaraldehyde, was comparable to the FITC conjugate. The peroxidase conjugate, prepared with periodate, was less effective.

scholarly journals SPECIFICITY IN THE COMBINATION OF FD FRAGMENTS WITH L CHAINS TO FORM HAPTEN-BINDING SITES

1966 ◽  
Vol 123 (5) ◽  
pp. 921-934 ◽  
Author(s):  
O. A. Roholt ◽  
G. Radzimski ◽  
D. Pressman
Keyword(s):  
Amino Acid ◽  
Propionic Acid ◽  
Binding Sites ◽  
Amino Acid Analysis ◽  
Binding Activity ◽  
Light Chains ◽  
Antibody Activity ◽  
Fc Fragment ◽  
Fab Fragments ◽  
Correct Alignment

In the work reported here we have shown that light chains and Fd fragments can be separated completely in propionic acid and then recombined to form Fab fragments with antibody activity. This experiment indicates that in the recombination a correct alignment of the Fd fragments and the L chains occurs to give a competent antibody site, just as occurs with the recombination of separated heavy and light chains of the antibody; thus the Fc fragment is not required for correct alignment. Fd fragments of antibody alone show very low binding activity toward the specific hapten. As is the case for the combination of heavy and light chains, the combination of Fd fragments and light chains also requires that both components come from antibody from the same rabbit in order to give binding sites. When they are derived from different rabbits producing antibody against the same antigen, they still give Fab fragments as shown by immunoelectrophoresis but do not have competent binding sites. An important observation is that the subunits of the papain digest fractions, FabI and FabII, have the capacity to cross-combine to form active Fab fragments with competent binding sites. FdI from FabI combines with LII chains from FabII to give the composite (FdI-LII) with good binding activity. Likewise, the composite (FdII-LI) has good binding activity. The composites from the two types of antibody molecules yielding different Fab fragments have antibody activity although heretofore these molecules have appeared to be different on the bases of chromatography and amino acid analysis. There is also a preferential combination of the Fd fragments to combine with the correct L fragments to give binding sites since this combination takes preference over the combination of Fd fragments of antibody with light chains of normal globulin (or of light chains of antibody with Fd fragments of normal globulin).

scholarly journals Anti-rabbit immunoglobulin G detection in complex medium by PM-RAIRS and QCM

2007 ◽  
Vol 22 (12) ◽  
pp. 2884-2890 ◽  
Author(s):  
Elisabeth Briand ◽  
Michèle Salmain ◽  
Chantal Compère ◽  
Claire-Marie Pradier
Keyword(s):  
Immunoglobulin G ◽  
Complex Medium ◽  
Rabbit Immunoglobulin

scholarly journals Differential reduction of interchain disulphide bonds of mouse immunoglobulin G

1968 ◽  
Vol 107 (6) ◽  
pp. 823-828 ◽  
Author(s):  
Alan R. Williamson ◽  
Brigitte A. Askonas
Keyword(s):  
Heavy Chain ◽  
Immunoglobulin G ◽  
Polyacrylamide Gel Electrophoresis ◽  
Partial Reduction ◽  
Free Light Chains ◽  
Myeloma Protein ◽  
Heavy Chains ◽  
Disulphide Bonds ◽  
Rabbit Immunoglobulin ◽  
Mouse Immunoglobulin

The relative lability of the interchain disulphide bonds of mouse G2a-myeloma protein 5563 was studied as a function of 2-mercaptoethanol concentration. Analysis of partial-reduction mixtures by polyacrylamide-gel electrophoresis and microdensitometry showed that the disulphide bonds between light and heavy chains are much more susceptible to reduction than the bonds between heavy chains. At a low concentration of 2-mercaptoethanol (10mm) the major dissociable products of mouse immunoglobulin G are heavy-chain dimers and free light chains. These findings contrast with the reported behaviour of rabbit immunoglobulin G, for which the lability of inter-heavy-chain bonds was found to exceed that of the bonds linking light and heavy chains (Hong & Nisonoff, 1965); the relative stability of rabbit immunoglobulin G interchain bonds was confirmed in the present study. Examination of human immunoglobulin G and an immunoglobulin G (γ2) of guinea pig showed that at least in the majority of molecules, as with mouse immunoglobulin G, the disulphide bonds between light and heavy chains are more susceptible to reduction than the inter-heavy-chain bonds.

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