Antigen-antibody crossed electrophoresis

Laurell, C.‐B.

doi:10.1016/0003-2697(65)90278-2

Cited by 1,369 publications

(324 citation statements)

References 2 publications

Supporting

Mentioning

309

Contrasting

Unclassified

Order By: Relevance

“…The method for performing CIE was based upon a modification (10) of Laurell's technique (11). Barbital-HCl buffer (ionic strength , = 0.02, pH 8.6) containing 1% (v/v) Triton X-100 was used throughout and incorporated into all gels.…”

Section: Materials and Methods Preparation Of Protoplasts And Membranmentioning

confidence: 99%

Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis.

Owen¹,

Saltón²

1975

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

on the surface. Of these antigens the major one has been identified as a succinylated mannan. Five of the principal immunoprecipitates unaffected by absorption with protoplasts were shown by zymograms to possess the following enzymic activities: succinate dehydrogenase (EC 1.3.99.1), ATPase (EC 3.6.1.3), NADH dehyrogenase (EC 1.6.99.3)(two separate components), and malate dehydrogenase (EC 1.1.1.37). These enzymes or enzyme-complexes are, therefore, not expressed on the outer surface of the protoplast membrane.Bacterial membranes perform many of the functions found in separate membranous organelles in the eukaryotic cell. Thus functions of electron transport, of active transport, and the biosynthesis of phospholipids and cell walls all occur in the bacterial membrane (1). In membranes of Gram-positive bacteria two principal regions, the plasma or cytoplasmic membrane and the mesosome, are recognizable (2) and these two entities have been isolated as homogeneous fractions from several species (3). Our investigations have been directed towards the elucidation of the structure-function relationships of the membranes of Micrococcus lysodeikticus by a multidisciplinary approach combining biochemical, immunochemical, and electron microscopic techniques. With conventional immunochemical methods, earlier studies in this laboratory showed the presence of three principal immunoprecipitates when membrane fractions of M. lysodeikticus reacted with membrane antiserum (4). These results did not reflect the anticipated complexity of the membrane, considering the multiplicity of the functions it performs and the number (about 30) of individual polypeptides found by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels (5, 6). Methods for the preparation of "total" membranes (i.e., plasma and mesosomal membranes), plasma membranes, and purified mannan from M. lysodeikticus are detailed elsewhere (5,8). Triton X-100 extracts of plasma membranes were obtained by treatment with detergent (final concentrations of 4%, w/v) for 1 hr at 220. Membrane residues (about 20%) sedimentable at 17,500 X g for 45 min were removed and the supernatant fractions (19 mg of protein per ml) were retained for analysis.Preparation of Antisera. Rabbits were immunized with "total" membranes (2 mg) given initially into both footpads and intradermally with Freund's complete adjuvant. Further injections of antigen without adjuvant were given weekly by subcutaneous injection at multiple sites. Subsequently, sera from four consecutive bleedings were pooled, and immunoglobulins were partially purified by precipitation with ammonium sulfate and dialysis against acetate buffer, pH 5.0 (9), and then concentrated by ultrafiltration to one-tenth of the original serum volume.Absorption of Anti-Membrane Serum with Protoplasts. Concentrated immunoglobulins (2.0 ml) were dialyzed against Buffer II (0.8 M sucrose, 10 mM Mg2+, 70 mM NaCl, 50 mM Tris'HCl, pH 8.6) and the final volume was adjusted to 6.0 ml. Aliquots (1.0 ml) were incubated for 1 hr at 22°with 0-3....

show abstract

Section: Materials and Methods Preparation Of Protoplasts And Membranmentioning

confidence: 99%

Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis.

Owen¹,

Saltón²

1975

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Conversion of C3 was assayed by antigen-antibody-crossed electrophoresis using anti-C3 in the second stage gel (17).…”

Section: Methodsmentioning

confidence: 99%

Role of Complement in Induction of Antibody Production in Vivo

Pepys

1974

The Journal of Experimental Medicine

312

108

View full text Add to dashboard Cite

There has been considerable recent progress in understanding of the mechanisms involved in cooperation between T and B lymphocytes in the induction of humoral antibody formation (see for example 1), but much still remains to be learned. The discovery of an interaction between lymphocytes and the fixed C3 component of complement (2-5), and the demonstration by Nussenzweig et al. that C3 receptors are a feature of B-cell populations (6-10), raise the possibility that C3 might play a part in this process (11). Germinal centers of lymphoid tissue, in which the induction of antibody production m a y occur, contain B cells, fixed C3, and complexes capable of fixing C3 (12). I have previously shown that treatment of mice in vivo with the C3-cleaving protein of cobra venom (CoF) 1 suppresses thymus-dependent but not thymus-independent antibody production, suggesting a possible role for C3 in lymphocyte cooperation (11).

show abstract

“…The concentration of HA in the inactivated virus vaccines prepared from different recombinant viruses was standardized by rocket immunoelectrophoresis (Laurell, 1965;Oxford, et al 1979), using monospecific antiserum to purified influenza virus RIT 4050 HA. This antiserum was prepared according to the methods of Brand & Skehel (1972).…”

Section: Rocket Immunoelectrophore8ismentioning

confidence: 99%

Antigenicity in hamsters of inactivated vaccines prepared from recombinant influenza viruses

Hamzawi

Jennings

Potter

1981

J. Hyg.

View full text Add to dashboard Cite

SummaryInactivated vaccines prepared from influenza virus strains obtained by the recombination of A/PR/8/34 (H1N1) or A/FM/1/47 (H1N1) viruses with A/Victoria/3/75 (H3N2) virus, were tested for their antigenicity in hamsters. The parental origin of the genes of each cloned recombinant virus was determined by polyacrylamide gel electrophoresis, and vaccines prepared from each strain by concentration, purification on sucrose density gradients and inactivation with formalin. All the recombinant strains used in these studies possessed surface haemagglutinin and neuraminidase antigens derived from the A/Victoria/75 parent strain.On inoculation into hamsters, at equivalent concentrations, these vaccines varied in their ability to induce haemagglutination-inhibiting (HI) antibodies in the serum. This variation was not dependent on concentration and was observed using neutralization and single radial haemolysis, as well as HI. The possible reasons for the findings are discussed.

show abstract

Antigen-antibody crossed electrophoresis

Cited by 1,369 publications

References 2 publications

Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis.

Antigenic and enzymatic architecture of Micrococcus lysodeikticus membranes established by crossed immunoelectrophoresis.

Role of Complement in Induction of Antibody Production in Vivo

Antigenicity in hamsters of inactivated vaccines prepared from recombinant influenza viruses

Contact Info

Product

Resources

About