title: Isolation and quantification of chitin-binding mistletoe lectin from mistletoe extracts and validation of this method.
abstract: 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.
authors.affiliation: Abteilung für Physikalische Biochemie, Physiologisch-chemisches Institut der Universität Tübingen, Tübingen, Germany.
authors.forename: Mirita
authors.initials: M
authors.lastname: Franz
authors.forename: Simone
authors.initials: S
authors.lastname: Vollmer
authors.forename: Roland
authors.initials: R
authors.lastname: Wacker
authors.forename: Sebastian
authors.initials: S
authors.lastname: Jäger
authors.forename: Rainer
authors.initials: R
authors.lastname: Scheer
authors.forename: Stanka
authors.initials: S
authors.lastname: Stoeva
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Voelter
mesh_terms: 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
chemical_list: D037102:Lectins; D010936:Plant Extracts; D002686:Chitin