Number of binding sites of rabbit macroglobulin antibody and its subunits

Onoue, Kaoru; Yagi, Yasuo; Grossberg, Allan L.; Pressman, David

doi:10.1016/0019-2791(65)90039-x

Cited by 107 publications

(34 citation statements)

References 47 publications

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“…Ra~ioimrnunoelectrophoresis (RIE).--Use of this method in the antilmpten antibody system was described previously (13,14).…”

Section: Fraaionation Of Normal Rabbitmentioning

confidence: 99%

“…Chromatography.--The same buffer systems as described previously for the separation of IgM and IgG antibody (14) were used for the separation of IgA antibody and IgG antibody.…”

Section: Diahylaminoethyl (Deae)-cdlulosementioning

confidence: 99%

“…It was prepared by reacting 30 moles of diazotized p-arsanilic acid per mole of RSA at pH 9. The The average number of Rp groups coupled to tyrosyl and histidyl residues of RSA was 7.8 moles per mole of RSA, as determined by spectrophotometry (20).Ra~ioimrnunoelectrophoresis (RIE).--Use of this method in the antilmpten antibody system was described previously (13,14). …”

mentioning

confidence: 99%

“…IgG and IgM antibodies were isolated and characterized for physical, chemical, and immunologic properties (14,15). Rabbit IgA antibody, which was found in only a few antisera, was shown to be similar to human IgA globulin in physical and antigenic properties relative to the other immunoglobulins.…”

mentioning

confidence: 99%

“…In Gel Filtration.--Gel filtration on Sephadex G-200 was carried out according to the method of Gelotte et al (18) using a Tris buffer of pH 8.0 containing 0.5 M NaC1 (14).…”

mentioning

confidence: 99%

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ISOLATION OF RABBIT IGA ANTIHAPTEN ANTIBODY AND DEMONSTRATION OF SKIN-SENSITIZING ACTIVITY IN HOMOLOGOUS SKIN

Onoue

Yagi

Pressman

1966

The Journal of Experimental Medicine

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Several classes of well characterized immunoglobulins are known in human and other animal species. Although antibodies of all classes of immunoglobulins combine with the antigen, they differ greatly in their activity in various immunologic phenomena which have requirements in addition to the combination of antigen and antibody.Only antibodies of a few immunoglobulin classes have been shown to be active in sensitizing skin. Furthermore, it has been reported that the antibody which sensitizes homologous skin is not necessarily the one which sensitizes heterologous skin. Thus, although human IgA antibody has been associated with reagin or antibody capable of sensitizing human skin (1-5), it did not sensitize guinea pig skin (6, 7). In contrast, human IgG antibody did not sensitize human skin but sensitized guinea pig skin. IgM antibody did not sensitize either skin (6, 7). Similar observations were made with guinea pig antibodies (8, 9) and with mouse antibodies (10-12).We have previously shown (13) that in rabbits antibody activity against simple haptenic groups is found in at least 3 classes of immunoglobulins, 7S T-globulin (IgG), 18S ~/1-or/~rmacroglobulin (IgM), and a/3t-globulin (IgA). IgG and IgM antibodies were isolated and characterized for physical, chemical, and immunologic properties (14, 15). Rabbit IgA antibody, which was found in only a few antisera, was shown to be similar to human IgA globulin in physical and antigenic properties relative to the other immunoglobulins. The results of gel filtration and nltracentrifugation indicated that rabbit IgA antibody is associated with a 9S globulin and possibly also with a 7S globutin (13).In the present study, the presence of 9S and 7S IgA antibody in rabbits was shown clearly by isolating them by gel filtration and chromatography from specificaUy purified antibody against the p-azobenzenearsonate group (Rp). In addition to physical and antigenic properties, skin-sensitizing activity in homologous (rabbits) and heterologous species (guinea pigs) was studied with 9S IgA, 7S IgA, IgM, and IgG antibodies. The results indicated that, in homologous species, the skin-sensitizing activity was shown by 7S IgA antibody, but not by 9S IgA antibody. The activity of IgG antibody was very low and IgM antibody did not sensitize rabbit skin. In contrast, IgG antibody was the most active in sensitizing skin of the heterologous species. Materials and MethodsAntisera.--Antisera against the p-azobenzenearsonate (Rp) group were obtained from rabbits immunized by intravenous injections of a p-azobenzenearsonate-bovine "/-globulin conjugate (16). Injections and bleedings were repeated weekly. Sera were collected during a period of 6 to 12 months, pooled, and used for the preparation of antibodies. Horse antiserum against rabbit globulin (horse anti-RG) (13), sheep antiserum against the whole macroglobulin fraction of normal rabbit serum (sheep anti-RMG) (14), and guinea pig antiserum against normal rabbit IgG (GP anti-IgG) (14) were the same antisera as those used in previous stu...

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“…Ra~ioimrnunoelectrophoresis (RIE).--Use of this method in the antilmpten antibody system was described previously (13,14).…”

Section: Fraaionation Of Normal Rabbitmentioning

confidence: 99%

“…Chromatography.--The same buffer systems as described previously for the separation of IgM and IgG antibody (14) were used for the separation of IgA antibody and IgG antibody.…”

Section: Diahylaminoethyl (Deae)-cdlulosementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

“…In Gel Filtration.--Gel filtration on Sephadex G-200 was carried out according to the method of Gelotte et al (18) using a Tris buffer of pH 8.0 containing 0.5 M NaC1 (14).…”

mentioning

confidence: 99%

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ISOLATION OF RABBIT IGA ANTIHAPTEN ANTIBODY AND DEMONSTRATION OF SKIN-SENSITIZING ACTIVITY IN HOMOLOGOUS SKIN

Onoue

Yagi

Pressman

1966

The Journal of Experimental Medicine

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Recognition ofE. coli tryptophan synthase by single-chain Fv fragments: comparison of PCR-cloning variants with the parental antibodies

et al. 1997

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The use of a recombinant antibody fragment instead of a complete antibody, as a conformational probe for protein structure and folding studies, can be technically advantageous provided that the recombinant fragment and its parental antibody recognize the antigen through the same mechanism. Monoclonal antibodies mAb19 and mAb93 are directed against the TrpB2 subunit of Escherichia coli tryptophan synthase and they have been extensively used as conformational probes of this protein. DNA sequences coding for single-chain variable fragments (scFv) of mAb19 and mAb93 were cloned and assembled by reverse transcription of the mRNAs from hybridomas and PCR amplification. A specialized plasmid vector, pFBX, was constructed; it enabled to express the scFvs as hybrids with the maltose-binding protein (MalE) in E. coli, and to purify them by affinity chromatography on cross-linked amylose. Six independent clones were sequenced for each hybridoma. All of them had differences in their nucleotide and amino acid sequences. A competition ELISA and the BIAcore biosensor apparatus were used to compare the energetics and kinetics with which the parental antibodies and the hybrids bound TrpB2. The antigen binding properties of the hybrids were close to those of the parental antibodies and they were only weakly affected by the differences of sequence between the clones, with one exception. The stability of one of the hybrids and its antigen binding properties were strongly modified by a change of Gln6 into Glu, introduced into its VH domain by the PCR primers. Simple models of bimolecular interaction did not fully account for the kinetic profiles obtained with the parental antibodies and the hybrids, and this complexity suggested the existence of a conformational heterogeneity in these molecules.

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Properties and modifications of a cryomacroglobulin possessing cold agglutinin activity

Deutsch

1969

Biopolymers

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synopsisThe physical properties of a pathological +-globulin with cold agglutinin activity and cryoglobulin solubility could be modified by changes in temperature and pH and upon dilution. The type of changes noted appear to simulate those expected in a readily dissociable antigen-antibody complex. Naturally occurring and chemically produced subunits of the +globulin and Fc-fragments of myeloma proteins diminished the cryoproperty of the +-globulin and effected changes in its physical properties but did not alter its cold agglutinin activity. Cryoglobulin properties could be conferred upon some purified +globulins by reacting them with Fc-fragments. Mercaptan dissociation of the cryomacroglobulin produces subunits with loss of the noted activities. Reaggregation of these subunits restores some of the native properties, but variable results which are dependent on the type of mercaptan employed are obtained. Attempts to explain the cold agglutinin activity of +globulin hybrids containing variable amounts of mercaptan-produced active and inert subunits suggest that activity requires two of the five subunits to be derived from the rM-cold agglutinin and to be adjacent to each other.

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Number of binding sites of rabbit macroglobulin antibody and its subunits

Cited by 107 publications

References 47 publications

ISOLATION OF RABBIT IGA ANTIHAPTEN ANTIBODY AND DEMONSTRATION OF SKIN-SENSITIZING ACTIVITY IN HOMOLOGOUS SKIN

ISOLATION OF RABBIT IGA ANTIHAPTEN ANTIBODY AND DEMONSTRATION OF SKIN-SENSITIZING ACTIVITY IN HOMOLOGOUS SKIN

Recognition ofE. coli tryptophan synthase by single-chain Fv fragments: comparison of PCR-cloning variants with the parental antibodies

Properties and modifications of a cryomacroglobulin possessing cold agglutinin activity

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