Production and Characterization of Monoclonal Antibodies Against Bacteroides forsythus and Wolinella recta

1988 ◽  
Vol 67 (3) ◽  
pp. 548-553 ◽  
Author(s):  
G. Werner-Felmayer ◽  
B. Guggenheim ◽  
R. Gmür
Keyword(s):  
Monoclonal Antibodies ◽  
Hybrid Cell ◽  
Subgingival Plaque ◽  
Direct Identification ◽  
Oral Flora ◽  
Myeloma Cells ◽  
Hybrid Cell Lines ◽  
Respective Species ◽  
Antigenic Heterogeneity

Hybrid cell lines producing monoclonal antibodies against Bacteroides forsythus or Wolinella recta were generated by fusion of SP2/0 or FO myeloma cells with splenocytes from Balb/c mice immunized with formalinized cells of B. forsythus FDC 331 or W. recta D13a-g, respectively. Enzyme-linked immunosorbent assays and indirect immunofluorescence tests were used to analyze the distribution of the recognized antigens on a panel of 70 strains representing 35 taxa, most of which are members of the oral flora. All monoclonal antibodies-eight against B. forsythus and six against W. recta-proved specific for the immunizing species. Four of the monoclonal antibodies against W. recta recognized antigens expressed by only some of the tested W. recta strains, thus confirming the earlier noted antigenic heterogeneity of this species. Antibody binding patterns consistent with those previously described or distinct for new serogroups could not, however, be observed. Four of the eight anti-B. forsythus monoclonal antibodies bound to only two of the three tested B. forsythus strains. All the remaining monoclonal antibodies detected every strain tested of the respective species against which they were raised. Preliminary results indicated that several of these antibodies should be very useful for the direct identification and quantification of these organisms in subgingival plaque.

scholarly journals Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies to Shiga-Like Toxins of I and II Types

2021 ◽  
pp. 83-88
Author(s):  
G. V. Kuklina ◽  
S. S. Ipatov ◽  
D. V. Pechenkin ◽  
A. V. Eremkin ◽  
A. A. Kytmanov ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Hybrid Cell ◽  
Sandwich Elisa ◽  
Minimum Detectable Concentration ◽  
Myeloma Cells ◽  
Hybrid Cell Lines

Objective – obtaining and characterization of hybrid cell lines producing monoclonal antibodies against I and II types of shiga-like toxins.Materials and methods. Shiga-like toxins obtained in “48thCentral Research Institute” of Ministry of Defense of Russian Federation (Kirov), BALB/c mice, myeloma cells SP2/0-Ag14 were used in research. Immune splenocytes and SP2/0-Ag14 myeloma cells were fused according to G. Kohler and C. Milstein method in De St. Fazekas and D. Scheidegger modifcation using 50 % polyethylene glycol. Hybrid cell lines producing specifc monoclonal antibodies were cloned by limited dilutions. Hybridomas growth and producing properties were studied in vitro and in vivo. Specifc activity of immune sera, culture and ascitic fluids were studied by indirect ELISA. Monoclonal antibodies from ascitic fluids were precipitated by saturated ammonium sulfate, followed by ion exchange chromatographyResults and discussion. 8 hybridomas producing monoclonal antibodies against I and II types shiga-like toxins were obtained. Hybridomas are characterized by stable proliferation and antibody-producing activity during 10 passages in vitro and 3 passages in vivo (observation period). Obtained monoclonal antibodies can be used for ELISA detection of I and II types shiga-like toxins. Minimum detectable concentration of shiga-like toxins in sandwich ELISA is 1 ng/ml. The possibility of detecting shiga-like toxins without typical differentiation was shown when using in the enzyme immunoassay a polyreceptor mixture of monoclonal antibodies for sensitizing the plate and a polyspecifc mixture of immunoperoxidase conjugates.

Construction and characterization of Chinese hamster cell EmtA EmtB double mutants

1983 ◽  
Vol 3 (5) ◽  
pp. 761-772
Author(s):  
S Chang ◽  
J J Wasmuth
Keyword(s):  
Ribosomal Protein ◽  
Cell Lines ◽  
Ribosomal Proteins ◽  
Hybrid Cell ◽  
Polyacrylamide Gel Electrophoresis ◽  
Chinese Hamster ◽  
Chinese Hamster Cell ◽  
Altered Form ◽  
Hybrid Cell Lines

Starting with hybrid cell lines between a Chinese hamster cell EmtA mutant and a Chinese hamster cell EmtB mutant, we have constructed cell lines that are homozygous for mutant alleles at both the emtA locus and the emtB locus, by using a two-step segregation protocol. The EmtA EmtB double mutants are approximately 10-fold more resistant to emetine inhibition than either of the parental mutants. Having both the EmtA mutation and the EmtB mutation expressed in the same cell also results in a level of resistance to cryptopleurine that is significantly higher than a simple additive effect of the two mutations alone. Analysis of ribosomal proteins by two-dimensional polyacrylamide gel electrophoresis demonstrated that a parental hybrid and a first-step segregant, which has lost the wild-type emtA allele, synthesize both a normal and an altered form of ribosomal protein S14, whereas an EmtA EmtB double mutant synthesizes only the altered form of this ribosomal protein. This result confirms that the emtB locus is the structural gene for ribosomal protein S14. Our results also suggest that the products of the emtA and emtB loci interact directly, indicating that the emtA locus, like the emtB locus, encodes a component of the ribosome.

Production and characterization of cytotoxic Thy-1 antibody-secreting hybrid cell lines Detection of T cell subsets

1979 ◽  
Vol 9 (11) ◽  
pp. 875-886 ◽  
Author(s):  
Phil Lake ◽  
Edward A. Clark ◽  
Manoocher Khorshidi ◽  
Geoffrey H. Sunshine
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Hybrid Cell ◽  
T Cell Subsets ◽  
Hybrid Cell Lines

Production and characterization of murine monoclonal antibodies against staphylococcal enterotoxins A and E

1991 ◽  
Vol 37 (8) ◽  
pp. 581-585 ◽  
Author(s):  
Kunihiro Shinagawa ◽  
Emiko Nishimura ◽  
Makoto Mitsumori ◽  
Naonori Matsusaka ◽  
Shunji Sugii
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibodies ◽  
The Other ◽  
Staphylococcal Enterotoxin A ◽  
Binding Assays ◽  
Spleen Cells ◽  
Myeloma Cells ◽  
Murine Monoclonal Antibodies ◽  
Competitive Binding Assays

Six murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin A (SEA) and enterotoxin E (SEE) were prepared by fusion of myeloma cells with mouse spleen cells immunized with SEA and SEE. Of five MAbs to SEA tested, two MAbs were reactive with only SEA, whereas three were specific for both SEA and SEE. On the other hand, one MAb to SEE was found to be specific for only SEE. To study specificities of the combining sites of these MAbs, competitive binding assays with either SEA or SEE and horseradish peroxidase conjugated MAbs were performed using unconjugated MAbs as inhibitors. The results obtained in the assays suggest that different epitopes may be located on SEA and that some of them may be cross-reacting epitopes between SEA and SEE. Key words: enterotoxins, monoclonal antibodies, Staphylococcus aureus.

Establishment of Hybrid Cell Lines Producing Monoclonal Antibodies to a Synthetic Peptide from the E1 Region of the Hepatitis C Virus

2007 ◽  
Vol 29 (1) ◽  
pp. 91-104 ◽  
Author(s):  
Ashraf A. Tabll ◽  
Samy B. Khalil ◽  
Reem M. El‐Shenawy ◽  
Gamal Esmat ◽  
Amr Helmy ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Lines ◽  
Synthetic Peptide ◽  
Hybrid Cell ◽  
Hybrid Cell Lines
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