Isolation and quantification of chitin-binding mistletoe lectin from mistletoe extracts and validation of this method.
A method was established to isolate and quantify small amounts of chitin-binding mistletoe lectin (cbML) from extracts of the mistletoe (Viscum album L.) by affinity and reverse phase high performance liquid chromatography. A validation, according to ICH guidelines, of this analytical method was carried out and showed that specificity, robustness and precision are guaranteed. In addition, linearity is ensured for a content between 0.6 and 4.1 microg/ml of cbML in the extracts and recovery was calculated to be in the range of 94 to 100%. So, accuracy of the method is guaranteed as well. As far as the range of the analytical method is concerned, a minimum of 1.2 microg and a maximum of 8.2 microg cbML can be incubated with the affinity material. Detection and quantitation limits were calculated to be 0.13 and 0.46 microg/ml cbML, respectively.
Franz
Mirita
M
Abteilung für Physikalische Biochemie, Physiologisch-chemisches Institut der Universität Tübingen, Tübingen, Germany.
Vollmer
Simone
S
Wacker
Roland
R
Jäger
Sebastian
S
Scheer
Rainer
R
Stoeva
Stanka
S
Voelter
Wolfgang
W
D000595:Amino Acid Sequence; D002138:Calibration; D002686:Chitin / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D037102:Lectins / Q000737:chemistry*; D014784:Mistletoe / Q000737:chemistry*; D008969:Molecular Sequence Data; D010936:Plant Extracts / Q000737:chemistry; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D037102:Lectins; D010936:Plant Extracts; D002686:Chitin