Method for measuring beta-lactam antibiotics

Patent metadata

Field	Value
Publication number	US-9689021-B2
Application number	US-201214350646-A
Country	US
Kind code	B2
Filing date	Oct 15, 2012
Priority date	Oct 14, 2011
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

The present invention relates to methods for determining the concentration of a free beta-lactam antibiotic in a biological sample. In particular, the present invention relates to methods for measuring a free beta-lactam antibiotic in a biological sample, comprising the steps of: (a) providing at least one class C beta-lactamase, a functional fragment or variant thereof; (b) providing at least one biological sample; (c) contacting said at least one class C beta-lactamase with said at least one biological sample; and (d) determining the concentration of said free beta-lactam antibiotic in said at least one biological sample.

First claim

The invention claimed is:

1. A method for determining concentration of a free beta-lactam antibiotic in a biological sample, comprising the steps of:
(a) providing at least one class C beta-lactamase comprising an amino acid sequence selected from the group consisting of SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10 and SEQ ID NO:12;
(b) providing at least one biological sample;
(c) contacting said at least one class C beta-lactamase with said at least one biological sample; and
(d) determining said concentration of said free beta-lactam antibiotic in said at least one biological sample.

2. A method for determining concentration of a free beta-lactam antibiotic in a biological sample, comprising the steps of:
(a) providing at least one class C beta-lactamase comprising an amino acid sequence of SEQ ID NO:8 or SEQ ID NO:12;
(b) providing at least one biological sample;
(c) contacting said at least one class C beta-lactamase with said at least one biological sample; and
(d) determining said concentration of said free beta-lactam antibiotic in said at least one biological sample.

3. The method according to claim 1 , wherein said biological sample is selected from the group comprising serum, blood, urine, interstitial fluid, saliva, tears, exudates, fluid collected from a deep tissue, fluid collected from a subcutaneous tissue.

4. The method of claim 3 , wherein said biological sample is serum.

5. The method according to claim 1 , wherein said biological sample is obtained from a subject receiving an antibiotherapy.

6. The method according to claim 1 , wherein said biological sample is obtained from a subject being treated for an infection caused by Gram positive and/or Gram negative micro-organisms.

7. The method according to claim 2 , wherein said biological sample is selected from the group comprising serum, blood, urine, interstitial fluid, saliva, tears, exudates, fluid collected from a deep tissue, fluid collected from a subcutaneous tissue, preferably said biological sample is serum.

8. The method of claim 7 , wherein said biological sample is serum.

9. The method according to claim 2 , wherein said biological sample is obtained from a subject receiving an antibiotherapy.

10. The method according to claim 2 , wherein said biological sample is obtained from a subject being treated for an infection caused by Gram positive and/or Gram negative micro-organisms.

Assignees

• Université de Liège
• Univ Catholique Louvain
• Université Libre de Bruxelles
• Université de Mons-Hainaut

Inventors

• Brans Alain
• Joris Bernard
• Delmarcelle Michael
• Marchand Jacqueline
• Tulkens Paul
• Hammaecher Catherine
• Goormaghtigh Erik
• De Coninck Joel

Classifications

• C12N9/86 Primary

  acting on amide bonds in cyclic amides, e.g. penicillinase {(3.5.2)} · CPC title

• C12Q1/34 Primary

  involving hydrolase · CPC title

• G01N2333/986

  acting on amide bonds in cyclic amides (3.5.2), e.g. beta-lactamase (penicillinase, 3.5.2.6), creatinine amidohydrolase (creatininase, EC 3.5.2.10), N-methylhydantoinase (3.5.2.6) · CPC title

• G01N33/9446

  Antibacterials · CPC title