Production of monoclonal antibodies to staphylococcal enterotoxin A

Edwin, Chitra; Tatini, S. R.; Strobel, Randy S.; Maheswaran, S. K.

doi:10.1128/aem.48.6.1171-1175.1984

Cited by 23 publications

(8 citation statements)

References 15 publications

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“…MAb. MAb were produced by growing hybridomas (C5, C3, B2II, B21) in tissue culture medium as described earlier (9). The supernatant from the tissue culture served as the MAb.…”

Section: Methodsmentioning

confidence: 99%

Specificity and cross-reactivity of staphylococcal enterotoxin A monoclonal antibodies with enterotoxins B, C1, D, and E

Edwin¹,

Tatini²,

Maheswaran³

1986

Appl Environ Microbiol

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The cross-reactivity of monoclonal antibodies produced against staphylococcal enterotoxin A with purified and crude enterotoxins B, C,, D, and E and the specificity of such reactions were evaluated by the indirect enzyme-linked immunosorbent assay and immunoblotting of Western blots (from sodium dodecyl sulfatepolyacrylamide gel electrophoresis) followed by autoradiography. Purified and crude enterotoxins B were also tested with polyclonal antibodies. Specificity of reactivity was demonstrated by immunoblotting of crude enterotoxin A, crude enterotoxin A treated with trypsin, crude enterotoxin E, and also with crude A, B, Cl, and D that were pretreated with Sepharose-4B-linked normal rabbit immunoglobulin G to remove protein A. A band corresponding to each staphylococcal enterotoxin was seen with monoclonal antibodies under all conditions tested and also with crude and purified enterotoxin B with two different (rabbit and goat) polyclonal antisera.

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“…MAb. MAb were produced by growing hybridomas (C5, C3, B2II, B21) in tissue culture medium as described earlier (9). The supernatant from the tissue culture served as the MAb.…”

Section: Methodsmentioning

confidence: 99%

Specificity and cross-reactivity of staphylococcal enterotoxin A monoclonal antibodies with enterotoxins B, C1, D, and E

Edwin¹,

Tatini²,

Maheswaran³

1986

Appl Environ Microbiol

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“…We have recently succeeded (6) in producing hybridomas which secreted monoclonal antibodies (MAb) to staphylococcal enterotoxin A (SEA). Tissue culture fluid from these clones exhibited high titers (106 to 107) in an indirect enzymelinked immunosorbent assay (ELISA).…”

mentioning

confidence: 99%

Nature and reactivity of staphylococcal enterotoxin A monoclonal antibodies

Edwin¹,

Tatini²,

Maheswaran³

1986

Appl Environ Microbiol

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Monoclonal antibodies from four clones (C5, C3, B2II, and B2I) directed against staphylococcal enterotoxin A were tested by the indirect enzyme-linked immunosorbent assay and double-gel immunodiffusion (micro-Ouchterlony) assay for the nature of heavy and light chain types. The reactivities of monoclonal antibodies were also tested by indirect enzyme-linked immunosorbent assay with various levels of purified staphylococcal enterotoxin A and various levels (dilutions) of monoclonal antibodies and saturation analysis-competitive indirect enzyme-linked immunosorbent assay. The heavy-chain isotype of monoclonal antibodies was found to be an unspecified subclass of immunoglobulin G1, and the light chain was the kappa type. Monoclonal antibodies from all of the clones exhibited high reactivity and nearly the same affinity to staphylococcal enterotoxin A in saturation analysis-competitive enzyme-linked immunosorbent assay. Purified immunoglobulin G from B2I yielded very high absorbance (1.2) at 405 nm with 1 ng of staphylococcal enterotoxin A as the coating antigen in the enzyme-linked immunosorbent assay. Monoclonal antibodies from B2I also neutralized the biological activity of staphylococcal enterotoxin A when tested by the kitten bioassay.

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“…The derived amino acid sequence of the 771-bp ORF of sezA+ had 77 and 79% sequence identities with SEA and SEE, respectively. It is reasonable to expect such closely related proteins to serologically cross-react, because mature forms of SEA and SEE, with 81% amino acid sequence identity, cross-react (15,28,43). The most likely reason that we were unable to demonstrate an enterotoxinlike sezA+ product is that the 771-bp ORF was not translated.…”

Section: Discussionmentioning

confidence: 90%

Identification of a bacteriophage containing a silent staphylococcal variant enterotoxin gene (sezA+)

1990

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A variant enterotoxin gene, referred to as sezA+, has been identified. Staphylococcus aureus FRI1106, a staphylococcal enterotoxin type D producer (Sed+), contained Hindlll fragments of 3.8 and 9.4 kilobase pairs (kbp) that hybridized in Southern blot analysis to a probe containing only staphylococcal enterotoxin type A structural gene sequences. Presumably, probe A-624 hybridized to the 9.4-kbp HindIll fragment because of the sequence homology between sea+ and sed+. This 9.4-kbp HindIII fragment, which was part of a staphylococcal plasmid, was isolated and ligated into an Escherichia coli plasmid vector; Sed+ E. coli recombinant clones were isolated. The 3.8-kbp HindUl fragment was shown to be part of a viable lysogenic bacteriophage, and it contained sezA+. This sezA4-containing fragment was cloned into E. coli, and its DNA sequence was determined. Examination of the nucleotide sequence revealed a 771-bp region that contained an open reading frame with 85 and 77% nucleotide and derived amino acid sequence identities with sea' and staphylococcal enterotoxin type A, respectively. This open reading frame has 83 to 50% nucleotide sequence identities with the other types of staphylococcal enterotoxin genes. sezA+ was shown to be transcribed into stable mRNA.However, the sezA' mRNA was not translated into an enterotoxinlike protein because it lacks an appropriate translation initiation codon. 1614 on August 3, 2020 by guest http://iai.asm.org/ Downloaded from

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Production of monoclonal antibodies to staphylococcal enterotoxin A

Cited by 23 publications

References 15 publications

Specificity and cross-reactivity of staphylococcal enterotoxin A monoclonal antibodies with enterotoxins B, C1, D, and E

Specificity and cross-reactivity of staphylococcal enterotoxin A monoclonal antibodies with enterotoxins B, C1, D, and E

Nature and reactivity of staphylococcal enterotoxin A monoclonal antibodies

Identification of a bacteriophage containing a silent staphylococcal variant enterotoxin gene (sezA+)

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