Monoclonal antibodies against species-specific cephalosporinase of Pseudomonas aeruginosa

Murakami, Kazuhisa; Yoshida, Tadashi

doi:10.1111/j.1432-1033.1985.tb08706.x

Cited by 7 publications

(6 citation statements)

References 27 publications

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“…These six neutralizing MAbs make up the group recognizing epitope B. We did not observe any significant enhancement of the enzyme activity as previously reported (9,32). Since the EIA-PADAC technique involves measuring the ,B-lactamase activity, it is surprising to find that the six neutralizing MAbs were positive by both EIAs.…”

Section: Methodsmentioning

confidence: 46%

Monoclonal antibodies to TEM-1 plasmid-mediated beta-lactamase

Morin

Patel

Levesque

et al. 1987

Antimicrob Agents Chemother

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At least 28 plasmid-mediated j-lactamases have been described in gram-negative bacteria. To assess the relationship among these enzymes, we produced and characterized 28 murine monoclonal antibodies to the TEM-1 plasmid-mediated ,i-lactamase. Radial immunodiffusion identified 3 monoclonal antibodies as immunoglobulin M (IgM), 18 as subclass IgGl, 2 as IgG2a, and 5 as IgG2b. Using a newly described enzyme immunoassay, cross-reactivity of 16 of these monoclonal antibodies was tested against 24 plasmid-determined ,-lactamases. The 16 monoclonal antibodies cross-reacted with TEM-2 and TLE-1 and, to a certain extent, SHV-1. Different levels of cross-reactivity were also observed with OXA-3 (11 of 16), OXA-7 (8 of 16), OXA-1 (2 of 16), OXA-6 (2 of 16), and AER-1 (2 of 16). Six monoclonal antibodies demonstrated partial neutralization of I8-lactamase activity. This study suggests that common epitopes are shared by nine biochemically distinct plasmid-mediated I-lactamases. On the basis of cross-reactivities with these monoclonal antibodies, we identified four epitopes on TEM-1, TEM-2, TLE-1, and SHV-1 P-lactamases.Four mechanisms of bacterial resistance to ,-lactam antibiotics have been described in gram-negative bacteria (13): (i) modification of cell-wall permeability (46), (ii) modification of penicillin-binding proteins (54), (iii) tolerance to the antibiotic (58), and (iv) enzymatic modification of the drug (34). Among these, the most frequent mechanism is the production of P-lactamases (EC 3.5.2.6) which hydrolyze the antibiotic, rendering it inactive (34). These enzymes are produced by almost all bacterial species and differ from each other in their enzymological and physicochemical properties. Although three classifications have been proposed (1, 42, 56), we tend to refer to ,B-lactamases as plasmid-mediated or chromosomal enzymes. At least 28 plasmid-mediated ,-lactamases are known (5,15,19,24,26,30,31,36,38,45,53), including the newly described BRO-1 (11), NPS-1 (25), OHIO-1 (51), and SAR-1 (39). Of all these enzymes, the most widely distributed is the TEM-1 P-lactamase (26,44,52).Three techniques are available to identify plasmidmediated P-lactamases precisely: analytical isoelectric focusing (27), DNA hybridization (23), and immunological assays (40,41). Several studies have reported immunological comparisons between plasmid-mediated P-lactamases using polyclonal antisera directed against TEM

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Section: Methodsmentioning

confidence: 46%

Monoclonal antibodies to TEM-1 plasmid-mediated beta-lactamase

Morin

Patel

Levesque

et al. 1987

Antimicrob Agents Chemother

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“…The substrate profiles were reported previously (10,11,17,25 diluted mixture was incubated at 30°C. Equal portions (0.2 ml) were periodically added to 2 ml of cephaloridine (110 ,uM) to measure the recovery of the enzyme activity.…”

mentioning

confidence: 99%

“…The other ,B-lactamases were partially purified as described elsewhere (9). Substrate specificities of the enzyme have been reported previously (10,11,17,25 …”

mentioning

confidence: 99%

“…Preparation of 1-lactamase. Cephalosporinase of Pseudomonas aeruiginosa SR24-12 and of Citrobacter freundii SR19 was purified by ion-exchange chromatography on carboxymethyl-Sephadex C-50 followed by gel filtration chromatography on Sephadex G-75 (10,11). The enzyme preparation of P. aeruginosa contained minor bands on isoelectric focusing (Fig.…”

mentioning

confidence: 99%

“…The other ,B-lactamases were partially purified as described elsewhere (9). Substrate specificities of the enzyme have been reported previously (10,11,17,25 (final concentration, 7.7 FiM) was incubated with the compound at various molar ratios in 0.5 ml of the buffer for the indicated time at 30°C. A 10-,ul portion was diluted with 5 ml of the buffer, and a 0.2-ml portion of the diluted mixture was added to 2 ml of cephaloridine (110 ,uM) as a substrate to measure the residual enzyme activity.…”

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confidence: 99%

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7 alpha-formylamino substituent confers beta-lactamase-inactivating potency on 1-oxacephalosporins

Murakami¹,

Doi²,

Yoshida³

1986

Antimicrob Agents Chemother

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acid (Fl) and 7ft-formylamino-7 -1 -2-[(4-ethyl-2,3-dioxopiperazine-1-yl)carbonylamino]-2-phenylacetamido}-3-[( 1-methyl-1H-tetrazol-5-yl)thiomethyl]-1-oxa-3-cephem-4-carboxylic acid (F2) were stable against penicillinases and, moreover, inactivated cephalosporinases of Pseudomonas aeruginosa, Citrobacterfreundii, and Enterobacter cloacae.Extensive studies of the inactivation of cephalosporinase of P. aeruginosa showed that it resulted from the formation of a transiently stable enzyme-compound complex. The 7a-formylamino substituent was involved in the enzyme inactivation, because 7a-methoxy congeners did not inactivate the enzyme. The number of compound molecules required for inhibition of an enzyme molecule was found to be 36 for Fl and 5.5 for F2, which suggests that the pathway to the complex formation branched off the hydrolysis pathway. Half-lives of the complexes were 400 min for Fl and 260 min for F2. 7a-Formylamino compounds Fl and F2 had antibacterial activities similar to those of 7fi-methoxy congeners against ,-lactamase-producing gram-negative bacteria, whereas they were less active against non-1-lactamase-producing strains of Escherichia coli and Staphylococcus aureus. The conclusion was that the 7a-formylamino substituent conferred the ability to inactivate cephalosporinase on the 1-oxacephalosporins tested, without much impairment of their antibacterial activity.,-Lactamase catalyzes cleavage of the amide bond of the P-lactam ring of ,-lactam antibiotics, rendering the antibiotics inactive, and r-lactamase production is the main cause of bacterial resistance to P-lactam antibiotics (16,21). Thus, the introduction of substituents which stabilize 3-lactam compounds against r-lactamases or inhibit the enzyme is very important for the development of antibiotics having high activities against f3-lactamase-producing bacteria.Cephalosporins containing the 7oc-methoxy substituent (cephamycins) were found in nature in the early 1970s (12,20). This substituent stabilizes cephamycins against 1-lactamases (3). Since then, 7ox-substituted cephalosporins and 6(-substituted penicillins have become a very important group of 1-lactam antibiotics. In screening programs, many 7(-and 7,B-substituted 1-oxacephalosporins have been synthesized, and their biological activities were tested in our laboratories (13). Recent discoveries of naturally occurring 7ot-formylaminocephalosporins in our and other laboratories (18,19,22) have stimulated the synthesis of two 7a-formylamino-1-oxacephalosporins in our laboratories. We examined the interaction of these two compounds with f-lactamases and found that the 7ot-formylamino-1-oxacephalosporins inactivated cephalosporinase by forming a transiently stable complex with the enzyme, but 7ot-methoxy congeners did not. (University of Manchester, Manchester, England), and E. coli ML1410 carrying R-plasmid RGN238 and Klebsiella pneumoniae GN69 were obtained from T. Sawai (Chiba University, Chibashi, Japan). Other strains were clinical isolates stocked in our laboratories. Preparation ...

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Immunochemical Prophylaxis against Pseudomonas aeruginosa

Collins

1993

Pseudomonas Aeruginosa as an Opportunistic Pathogen

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Monoclonal antibodies against species-specific cephalosporinase of Pseudomonas aeruginosa

Cited by 7 publications

References 27 publications

Monoclonal antibodies to TEM-1 plasmid-mediated beta-lactamase

Monoclonal antibodies to TEM-1 plasmid-mediated beta-lactamase

7 alpha-formylamino substituent confers beta-lactamase-inactivating potency on 1-oxacephalosporins

Immunochemical Prophylaxis against Pseudomonas aeruginosa

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