Skip to main content
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1974 Dec;6(6):676–679. doi: 10.1128/aac.6.6.676

Immunogical Study of Anti-Beta-Lactamase Antibodies by Acidimetric Methods

François Le Goffic 1,2, Jane Andrillon-Spiegel 1,2, Robert Letarte 1,2
PMCID: PMC444718  PMID: 4217581

Abstract

A beta-lactamase was extracted from an Escherichia coli K-12 strain carrying the R-TEM plasmid and has been purified by affinity chromatography. Antisera to this enzyme were prepared in the rabbit, and the enzyme-antibody neutralization reaction has been evaluated with acidimetric methods (pH stat or pH meter). Under defined experimental conditions, it is now possible to clearly illustrate the enzyme-antibody reaction by means of accurate, rapid, and simple-to-perform methods. These methods are in accord with the specificity of the two reactive partners and allow the detection of the enzyme in a crude bacterial extract which would eventually contain more than one beta-lactamase.

Full text

PDF
676

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Dale J. W., Smith J. T. A direct comparison of two unusual R-factor-mediated -lactamases. Biochem J. 1972 Jun;128(1):173–174. doi: 10.1042/bj1280173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dale J. W., Smith J. T. Some relationships between R-factor and chromosomal -lactamase in Gram-negative bacteria. Biochem J. 1971 Jul;123(4):507–512. doi: 10.1042/bj1230507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Datta N., Richmond M. H. The purification and properties of a penicillinase whose synthesis is mediated by an R-factor in Escherichia coli. Biochem J. 1966 Jan;98(1):204–209. doi: 10.1042/bj0980204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Jack G. W., Richmond M. H. A comparative study of eight distinct beta-lactamases synthesized by gram-negative bacteria. J Gen Microbiol. 1970 Apr;61(1):43–61. doi: 10.1099/00221287-61-1-43. [DOI] [PubMed] [Google Scholar]
  5. Kazmierczak A., Philippon A., Chardon H., Labia R., Le Goffic F. Constantes enzymatiques (Km et Vmax) des beta-lactamases mesurées par une méthode micro-acidimétrique couplée a l'ordinateur. Ann Microbiol (Paris) 1973 Oct;124(3):259–268. [PubMed] [Google Scholar]
  6. Labia R., Andrillon J., Le Goffic F. Computerized microacidimetric determination of beta lactamase Michaelis-Menten constants. FEBS Lett. 1973 Jun 15;33(1):42–44. doi: 10.1016/0014-5793(73)80154-1. [DOI] [PubMed] [Google Scholar]
  7. Linström E. B., Boman H. G., Steele B. B. Resistance of Escherichia coli to penicillins. VI. Purification and characterization of the chromosomally mediated penicillinase present in ampA-containing strains. J Bacteriol. 1970 Jan;101(1):218–231. doi: 10.1128/jb.101.1.218-231.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ogawara H., Maeda K., Umezawa H. A -lactamase of Escherichia coli. Biochim Biophys Acta. 1972 Nov 10;289(1):203–211. doi: 10.1016/0005-2744(72)90123-4. [DOI] [PubMed] [Google Scholar]
  9. POLLOCK M. R. An immunological study of the constitutive and the penicillin-induced penicillinases of Bacillus cereus, based on specific enzyme neutralization by antibody. J Gen Microbiol. 1956 Feb;14(1):90–108. doi: 10.1099/00221287-14-1-90. [DOI] [PubMed] [Google Scholar]
  10. POLLOCK M. R. The cell-bound penicillinase of Bacillus cereus. J Gen Microbiol. 1956 Aug;15(1):154–169. doi: 10.1099/00221287-15-1-154. [DOI] [PubMed] [Google Scholar]
  11. Pitton J. S. Mechanisms of bacterial resistance to antibiotics. Ergeb Physiol. 1972;65:15–93. doi: 10.1007/3-540-05814-1_2. [DOI] [PubMed] [Google Scholar]
  12. RICHMOND M. H. Immunological properties of exopenicillinase synthesized by Bacillus cereus 569/H in the presence of amino acid analogues. Biochem J. 1960 Oct;77:112–121. doi: 10.1042/bj0770112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Richmond M. H., Sykes R. B. The beta-lactamases of gram-negative bacteria and their possible physiological role. Adv Microb Physiol. 1973;9:31–88. doi: 10.1016/s0065-2911(08)60376-8. [DOI] [PubMed] [Google Scholar]
  14. Roupas A., Pitton J. S. R factor-mediated and chromosomal resistance to ampicillin in Escherichia coli. Antimicrob Agents Chemother. 1974 Feb;5(2):186–191. doi: 10.1128/aac.5.2.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Yamagishi S., O'Hara K., Sawai T., Mitsuhashi S. The purification and properties of penicillin beta-lactamases mediated by transmissible R factors in Escherichia coli. J Biochem. 1969 Jul;66(1):11–20. doi: 10.1093/oxfordjournals.jbchem.a129111. [DOI] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES