scholarly journals A STUDY OF IMMUNOGLOBULIN STRUCTURE

1969 ◽  
Vol 130 (2) ◽  
pp. 401-415 ◽  
Author(s):  
R. Quattrocchi ◽  
D. Cioli ◽  
C. Baglioni
Keyword(s):  
Light Chain ◽  
Peptide Mapping ◽  
Gel Filtration ◽  
Antigenic Determinants ◽  
Minimal Size ◽  
Bence Jones Protein ◽  
Peptide Map ◽  
Peptide Maps ◽  
Similar Peptide ◽  
Bence Jones Proteins

102 human Bence Jones proteins have been purified by gel filtration, digested with trypsin, and analyzed by peptide mapping. In several cases Bence Jones "fragments", corresponding to the variable half of the corresponding proteins, were observed. The peptide maps of the proteins were compared to establish whether any identical proteins were present in the sample analyzed. No Bence Jones protein showed a peptide map identical to that of any other protein, although remarkable similarities in the peptide maps were observed for some proteins. Two proteins that gave very similar peptide maps were then examined in detail, by purifying and analyzing the tryptic peptides. It was then found that these two proteins differ in amino acid sequence in at least six positions. The probability of not finding two identical sequences by examining a sample extracted from populations of light chains of different sizes has been calculated. This has led to an estimate of the minimal size of the population of light chain sequences in humans. The number of light chain sequences appears to be at least a few thousand. Information on the frequency of Inv and Oz antigenic determinants and on the relative frequency of subtypes of K chains has been obtained. Proteins of KI subtype are found most frequently. The possibility that different subtypes may be predominant in different species is discussed in relation to the evolutionary arguments used in favor of the somatic theories on the origin of variability of immunoglobulin chains.

scholarly journals A STUDY OF IMMUNOGLOBULIN STRUCTURE

1966 ◽  
Vol 124 (3) ◽  
pp. 307-330 ◽  
Author(s):  
C. Baglioni ◽  
D. Cioli
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Peptide Mapping ◽  
Gel Filtration ◽  
Bence Jones Protein ◽  
Type K ◽  
The Common ◽  
L Proteins ◽  
Peptide Maps ◽  
Bence Jones Proteins

Urinary proteins of patients with myeloma, prepared by precipitation with ammonium sulphate, have been separated by gel filtration on Sephadex G-100 after reduction and aminoethylation. Many specimens separated into a major peak of Bence Jones protein and into minor peaks of albumin, a protein tentatively identified with heavy chain and a smaller molecular weight protein corresponding to the variable portion of the corresponding Bence Jones protein. The Bence Jones protein purified by gel filtration was analyzed by electrophoresis and by peptide mapping after tryptic digestion. The peptide maps of 24 type K and 20 type L Bence Jones proteins were compared. A set of common peptides was identified in the peptide maps of the Bence Jones proteins of the same type; the common peptides of type K proteins were completely different from the common peptides of type L proteins. The patterns of distinctive peptides was compared; no similarities were found between distinctive peptides of type K and of type L proteins. Some similarities were observed in the distinctive peptides of proteins of the same type. The similarities involved in many cases peptides containing cysteine, whereas similarities in other peptides were limited. This observation suggested that the amino acid sequence around the cysteines of the variable NH2-terminal half of the Bence Jones proteins may show less variability than other sequences. A few proteins of the same type differed in all their distinctive peptides, an indication that multiple amino acid differences exist between individual Bence Jones proteins. The genetic mechanisms responsible for the variability in the amino acid sequence of the NH2-terminal half of the light chains of immunoglobulins are discussed in view of the results of the comparison by peptide mapping of the Bence Jones proteins.

scholarly journals CATABOLIC ORIGIN OF A BENCE JONES PROTEIN FRAGMENT

1968 ◽  
Vol 128 (3) ◽  
pp. 517-532 ◽  
Author(s):  
D. Cioli ◽  
C. Baglioni
Keyword(s):  
Amino Acids ◽  
Free Amino Acids ◽  
Free Amino ◽  
Peptide Mapping ◽  
Gel Filtration ◽  
Urinary Proteins ◽  
Small Peptides ◽  
Protein Fragment ◽  
Bence Jones Protein ◽  
Bence Jones Proteins

Gel filtration analysis of the urinary proteins of some patients with myeloma has shown the presence of "fragments" of Bence Jones proteins which correspond to the variable half of these proteins. Experiments have been carried out to establish the origin of a "fragment" observed in a patient who excreted a large amount of this protein. Labeled homologous Bence Jones protein has been injected into this and other control patients. Excretion of labeled "fragment" has been observed in all. Analysis by peptide mapping and radio-autography of this labeled "fragment" isolated from the urine showed that the invariable half of the Bence Jones protein was not excreted; it seemed thus likely that the invariable half was metabolized to small peptides and free amino acids. A labeled Bence Jones protein which was excreted without any accompanying "fragment" was injected into the patient who excreted large amounts of "fragment." No excretion of labeled "fragment" was observed. It was thus concluded that the property of being degraded to "fragment" is characteristic of some "fragile" Bence Jones proteins and is not determined by the patient. Incubation with serum or urine of the "fragile" Bence Jones protein failed to produce any "fragment." "Fragments" of Bence Jones proteins are thus most likely formed during excretion of these proteins through the kidney and are products of the catabolism of Bence Jones proteins.

Historical perspectives in clinical pathology: Bence Jones protein—early urine chemistry and the impact on modern day diagnostics

2020 ◽  
pp. jclinpath-2020-206675
Author(s):  
Sheromna Sewpersad ◽  
Tahir S Pillay
Keyword(s):  
Light Chain ◽  
Clinical Pathology ◽  
Light Chains ◽  
Immunoglobulin Light Chains ◽  
Bence Jones Protein ◽  
Historical Perspectives ◽  
Serum Free Light Chain ◽  
The Impact ◽  
Made In ◽  
Bence Jones Proteins

This is the third in the series of historical articles dealing with developments in clinical pathology. Bence Jones proteins are immunoglobulin light chains found in excessive quantities in urine in multiple myeloma and are believed to be one of the first tumour markers ever discovered . Dr Henry Bence Jones is credited with the discovery of this protein in 1847 that bears his name and he can also be regarded as the first chemical pathologist/clinical chemist. Since then, numerous advances and refinements have been made in the measurement and detection of urine light chain proteins which have resulted in the current sensitive serum free light chain assays used today.

scholarly journals BENCE JONES PROTEINS AND LIGHT CHAINS OF IMMUNOGLOBULINS

1969 ◽  
Vol 130 (6) ◽  
pp. 1295-1311 ◽  
Author(s):  
Alan Solomon ◽  
Carla L. McLaughlin
Keyword(s):  
Light Chain ◽  
Light Chains ◽  
Bence Jones Protein ◽  
Amino Terminal ◽  
Type K ◽  
Myeloma Proteins ◽  
Immunoglobulin Molecule ◽  
Polypeptide Chains ◽  
Bence Jones Proteins

Three distinct classes of κ light polypeptide chains have been detected immunochemically by an antiserum (R185) prepared against a κ Bence Jones protein with a glutamyl amino terminal residue. This antiserum had specificity for κ light chains with glutamyl amino terminal residues and differentiated κ-chains with aspartyl amino terminal residues into two classes: the three κ-chain classes have been designated as κglu, κaspII, and κaspI. The ability of antiserum R185 to detect these antigenic differences on the intact immunoglobulin molecule, as well as on the isolated light chain or Bence Jones protein, made feasible the direct classification of type K myeloma proteins and M-macroglobulins (Waldenström). The multispecificity of the antiserum permitted the quantitation of type κglu light chains in normal, hypergammaglobulinemic, and hypogammaglobulinemic sera. Whereas the distribution of myeloma proteins and Bence Jones proteins in the κglu class correlated with the distribution of κglu chains in normal and hypergammaglobulinemic sera, the M-macroglobulins in the κglu class represented 90% of the total M-macroglobulins tested and revealed a marked divergence from the range of 24–31% of κglu immunoglobulins in normal sera. A preponderance of κglu chains was detected in the sera from patients with non-sex-linked hypogammaglobulinemia and represented 60–77% of the total type K light chain content. The controlled cleavage of a Bence Jones protein representative of each κ-chain class into its variant half and constant half made possible the localization on the light polypeptide chain, the reactive sites for which antiserum R185 had specificity. The correlations between immunochemical and structural classification of κ light chains are discussed.

Interference of polyclonal free light chains with identification of Bence Jones proteins

1993 ◽  
Vol 39 (8) ◽  
pp. 1734-1738 ◽  
Author(s):  
P P Hess ◽  
W Mastropaolo ◽  
G D Thompson ◽  
S S Levinson
Keyword(s):  
Gel Filtration ◽  
Double Diffusion ◽  
Light Chains ◽  
Gel Chromatography ◽  
Bence Jones Protein ◽  
Free Light Chains ◽  
Isoelectric Points ◽  
Immunofixation Electrophoresis ◽  
Sample Dilution ◽  
Bence Jones Proteins

Abstract We present a case in which kappa free light chains caused difficulty in interpreting classical urinary immunoelectrophoresis, but immunofixation electrophoresis (IFE) demonstrated the presence of a lambda-Bence Jones protein. Analysis of the urine by Ouchterlony double diffusion and IFE after gel-filtration chromatography showed that the difficulty was caused by the presence of large amounts of polyclonal free light chains. The workup also demonstrated that although IFE is the more sensitive and specific technique, IFE performed on concentrated urinary samples is especially subject to misinterpretation unless densely staining patterns are diluted and reassayed. This process of sample dilution provides a means for titrating antigen and antibody concentrations such that condition-specific patterns become visible on the gel. This workup also shows that, at some dilutions, polyclonal free light chains may migrate in the same manner as an oligoclonal band in a so-called ladder configuration. These bands were observed from both monomeric and dimeric fractions isolated by gel chromatography, consistent with reports that this pattern is largely linked to the isoelectric points of the molecules. We speculate that, in rare instances, the distinction between polyclonal and monoclonal kappa free light chains migrating as a ladder-banding pattern may be equivocal.

scholarly journals ANTIGENIC RELATIONSHIPS OF BENCE JONES PROTEINS, MYELOMA GLOBULINS, AND NORMAL HUMAN γ-GLOBULIN

1963 ◽  
Vol 117 (1) ◽  
pp. 81-104 ◽  
Author(s):  
Shunsuke Migita ◽  
Frank W. Putnam
Keyword(s):  
Type A ◽  
Antigenic Determinants ◽  
Type I ◽  
Starch Gel Electrophoresis ◽  
Type Ii ◽  
Type B ◽  
Bence Jones Protein ◽  
Starch Gel ◽  
Antigenic Relationships ◽  
Bence Jones Proteins

By means of immunodiffusion and immunoelectrophoresis study has been made of antigenic relationships of Bence Jones proteins, and the three classes of normal and pathological immunoglobulins, 7S γ, ß2A, and ß2M. All thirty-nine Bence Jones proteins studied could be classified into either one of two distinct antigenic types, A or B. Both types are related to the immunoelectrophoretically slow (S) fragment of a papain digest of normal γ-globulin; B is related more closely than A, but neither has antigenic determinants in common with the fast (F) fragment. The 7S γ myeloma globulins were either immunological type I or II. The papain digests of these proteins produced the S and F precipitin lines in immunoelectrophoresis but multiple bands in starch gel electrophoresis, especially in the F region. The S fraction of type I myeloma globulins is antigenically similar to Bence Jones protein of type B, and the S component of type II myeloma globulins has antigenic determinants in common with type A Bence Jones protein. Correspondingly, myeloma patients with type I globulins and proteinuria usually excrete type B Bence Jones proteins, whereas patients with type II excrete type A proteins. The F fragment is the part common to normal 7S γ-globulin and types I and II myeloma globulins but is absent in ß2A and ß2M pathological globulins and in both types of Bence Jones proteins. Papain digests of ß2A myeloma globulins produced a single precipitin line in immunoelectrophoresis. ß2A myeloma globulins appeared to have two antigenic units, one in common with type B Bence Jones protein and normal γ-globulin, and another specific to ß2A. The ß2A myeloma patients excreted type B Bence Jones protein. The papain digest of a macroglobulin produced two precipitin lines, the faster of which had antigenic determinants in common with type B Bence Jones protein, the slower seemed specific for the macroglobulin. Five serum micromolecular globulins proved to be either type A or B Bence Jones proteins. From the above results, an antigenic map was constructed showing which determinants are shared and which are specific for normal 7S γ-globulin, types I and II myeloma globulins, ß2A myeloma globulins, a macroglobulin, and types A and B Bence Jones proteins.

Synthesis of hepatitis B surface antigen pre-S2 region fragments and studies on their immunogenicity

1988 ◽  
Vol 53 (11) ◽  
pp. 2952-2956 ◽  
Author(s):  
Bernard Lammek ◽  
Zbigniew Maćkiewicz ◽  
Izabela Derdowska ◽  
Hanna Świderska ◽  
Adam Nowosławski ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Surface Antigen ◽  
Solid Phase ◽  
Hepatitis B Surface Antigen ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Antigenic Determinants ◽  
Phase Method ◽  
Peptide Fragments ◽  
Humoral Immune

Two peptide fragments of hepatitis B surface antigen pre-S2 region were synthesized by the solid phase method. The peptides were purified by gel filtration or ion-exchange chromatography on Sephadex SP-C-25. Both peptides induced a cellular and humoral immune response in rabbits. The results showed that fragment 14-22 of pre-S2 region contains one of the antigenic determinants.

scholarly journals A second rabbit kappa isotype.

1982 ◽  
Vol 156 (2) ◽  
pp. 585-595 ◽  
Author(s):  
A Benammar ◽  
P A Cazenave
Keyword(s):  
Light Chain ◽  
Immunoglobulin G ◽  
Antigenic Determinants ◽  
Gene Encoding ◽  
Genetic Studies ◽  
B Locus ◽  
Different Populations ◽  
Domestic Rabbits ◽  
Chain Type

Immunoglobulin G (IgG) from the rabbit strain Basilea was previously shown to contain two distinct populations of molecules one with light chain belonging to the known lambda isotype and the others to a new kappa-like L chain type. Alloantisera prepared against the Basilea IgG are directed against the kappa-like light chain (anti-bas antisera). All Basilea rabbits express kappa-like chains recognized by anti-bas sera, but IgG from other domestic rabbits did not react with these antisera. Genetic studies of wild rabbits belonging to different populations show that the bas+ phenotype could be found in heterozygous rabbits as well as those homozygous at the b locus. The gene encoding the bas+ light chain is closely linked to the b locus. Moreover, antigenic determinants recognized by anti-bas antibodies and antigenic determinants recognized by antibodies directed against allotypic determinants of the b series are located on distinct IgG molecules. These results show that there are two rabbit kappa isotypes: the kappa 1 isotype, bearing allotypic determinants of the b series, and the kappa 2 isotype, for which bas+ chain is one of the allotypic forms. The kappa 1 and kappa 2 isotypes are controlled by closely linked genes.

Immunochemical Characterization of a Factor IX Inhibitor Following Anamnestic Response

1975 ◽  
Author(s):  
H. R. Roberts ◽  
C. R. Fuller ◽  
H. Worden ◽  
J. Stuart ◽  
H. Reisner ◽  
...  
Keyword(s):  
Light Chain ◽  
Gel Filtration ◽  
Complete Removal ◽  
Igg Subclasses ◽  
Factor Ix ◽  
Biologically Active ◽  
Anamnestic Response ◽  
Polyclonal Igg ◽  
Monospecific Antisera ◽  
A Minor

We previously characterized a human inhibitor for Factor IX in patient P.W.B, with Hemophilia B as an IgG4, λ immunoglobulin of restricted electrophoretic mobility. This restriction to a minor IgG subclass led us to characterize a second Factor IX inhibitor occurring in patient R. J. after an anamnestic response to Factor IX. On preparative zone electrophoresis the inhibitor migrated with a broad zone of mobility in the anodal portion of the γ peak and was restricted to the anodal portion of the IgG containing fractions. Gel filtration on calibrated 1.5 M sepharose columns revealed inhibitor activity in fractions corresponding to a molecular weight of 150,000. The inhibitor was further characterized by the technique of antibody neutralization using monospecific antisera to immunoglobulin classes, subclasses and light chain types in the zone of antibody excess. The inhibitor was completely neutralized by antibody to IgG whereas antisera to IgA, IgM, IgD and IgE had no effect. Neutralization was abolished by absorption of the IgG antiserum with purified IgG. Neutralization with antisera specific for light chains indicated a mixture of light chain types with an estimated ϰ/λ ratio of 6/1. Neutralization with antisera specific for IgG subclasses revealed a mixture of IgG subclasses. The Factor IX inhibitor was thus characterized as a polyclonal IgG immunoglobulin. Sepharose conjugates of R. J. globulin effect complete removal of Factor IX from normal plasma on an immunoabsorbent column and biologically active Factor IX may be eluted with 1600-fold purification.

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