XXII. BIOCHEMICKÝ ZJAZD - Jesseniova lekárska fakulta

Posters
58.
DEVELOPMENT OF a DETECTION TOOL TO FOLLOW THE SPECIFIC
ACTIVITY OF BUTYRYLCHOLINESTEraSE IN HUMAN PATIENTS
Katarína Mrvová and Anna Hrabovská
Dpt. of pharmacology and toxicology, Faculty of Pharmacy, Comenius University,
Odbojárov 10, 832 32 Bratislava
Many hypotheses have been proposed in afford to explain a butyrylcholinesterase (BChE)
inter-individual variability in humans, e.g., different expression levels; different catalytic
properties. Yet, it has been impossible to study this topic due to the lack of an efficient
detection tool. However, we have recently generated a selective and specific antibody
against human BChE which allows us to address this problem. The aim of the project
was to develop an ELISA assay for detection of BChE activity and use this tool to study
the inter-individual BChE activity variation in humans.
Human plasma was prepared from the capillary blood of 86 healthy human volunteers
(age 20 – 23; BMI = 17,6-28,4). The Ellman’s assay was used at the conditions determined
in our laboratory (see the abstract of D. Neuschlova). ELISA has been used to determine
specific activity. All samples were tested in triplicates.
In the first step we determined the conditions for the ELISA assay. The lowest saturating
dilution of the primary antibody and the experimental serum dilution were determined
from the saturation curves. In the second step, human sera were tested for the total
BChE activity (Ellman’s assay) and for the specific BChE activity in excess plasma (ELISA)
and compared. Our results suggest that an inter-individual BChE activity in humans is
a caused by both, higher expression level and different catalytic properties.
Acknowledgement: The project was supported by APVV grant SK-CZ-0028-09.
XXII. Biochemistry Congress, Martin
179