Monoclonal antibodies that identify gram-negative bacteria using the magnetic immunoluminescence assay

Thirteen murine hybridomas capable of producing monoclonal antibodies to somatic antigens on Bradyrhizobium japonicum were developed and an indirect enzyme-linked immunosorbent assay was used to test reactivity of the antibodies against 20 strains of B. japonicum. Although polyclonal antisera from mice immunized with strains of B. japonicum reacted with bacterial cells of all 20 strains, individual monoclonals were more specific. Some antibodies reacted with as few as 2 and one with as many as 11 strains. On the basis of reactivity with the set of 13 monoclonal antibodies, the 20 strains of B. japonicum could be divided arbitrarily into five groups. Three of five monoclonal antibodies tested reacted with bacteroids taken directly from soybean nodules. One monoclonal bound to cells of five species of Rhizobium, but none of the 13 reacted with gram-negative bacteria representing six other genera. Treatment of cells with reagents and heat indicated the chemical nature of the antigens to five of the monoclonals. Antigen reactive with one antibody was destroyed by periodate oxidation indicating that it was a polysaccharide. Two antigens were probably proteins as they could be digested by trypsin and denatured by heat. Two others were inactivated by all three treatments suggesting they were glycoproteins.

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Shared idiotypes among monoclonal antibodies specific for different immunodominant sugars of lipopolysaccharide of different Gram-negative bacteria.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.79.5.1616 ◽

1982 ◽

Vol 79 (5) ◽

pp. 1616-1620 ◽

Cited By ~ 21

Author(s):

J. Hiernaux ◽

C. A. Bona

Keyword(s):

Monoclonal Antibodies ◽

Gram Negative Bacteria ◽

Gram Negative

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Epitope mapping of monoclonal antibodies specific for the directly cross-linkedmeso-diaminopimelic acid peptidoglycan found in the anaerobic beer spoilage bacteriumPectinatus cerevisiiphilus

Canadian Journal of Microbiology ◽

10.1139/w99-071 ◽

1999 ◽

Vol 45 (9) ◽

pp. 779-785 ◽

Cited By ~ 11

Author(s):

Barry Ziola ◽

Sheryl L Gares ◽

Brandene Lorrain ◽

Lori Gee ◽

W M Ingledew ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Outer Membrane ◽

Epitope Mapping ◽

Sodium Dodecyl ◽

Diaminopimelic Acid ◽

Gram Negative Bacteria ◽

Gram Negative ◽

Spoilage Bacteria ◽

Beer Spoilage ◽

Bacterial Outer Membrane

Nineteen monoclonal antibodies (Mabs) were isolated based on reactivity with disrupted Pectinatus cerevisiiphilus cells. All of the Mabs reacted with cells from which the outer membrane had been stripped by incubation with sodium dodecyl sulphate, suggesting the peptidoglycan (PG) layer was involved in binding. Mab reactivity with purified PG confirmed this. Epitope mapping revealed the Mabs in total recognize four binding sites on the PG. Mabs specific for each of the four sites also bound strongly to disrupted Pectinatus frisingensis, Selenomonas lacticifix, Zymophilus paucivorans, and Zymophilus raffinosivorans cells, but weakly to disrupted Megasphaera cerevisiae cells. No antibody reactivity was seen with disrupted cells of 11 other species of Gram-negative bacteria. These results confirm that a common PG structure is used by several species of anaerobic Gram-negative beer spoilage bacteria. These results also indicate that PG-specific Mabs can be used to rapidly detect a range of anaerobic Gram-negative beer spoilage bacteria, provided the bacterial outer membrane is first removed to allow antibody binding.Key words: beer spoilage, epitope mapping, monoclonal antibodies, Pectinatus, peptidoglycan.

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